Chromosome Based Strategies to Decipher the Structure and Evolution of the Hexaploid Wheat Genome: Chromosome 3B,a Case Study

With 17% of all crop area, wheat is the staple food for 40% of the world’s population. Improvement in bread wheat quality and yield in the context of sustainable agriculture is needed in the next decades to meet human

Genetic Structure of the Oriental River Prawn (Macrobrachium nipponense) from the Yangtze and Lancang Rivers, Inferred from COI Gene Sequence

This study analyzed nueleotide sequences from the mitochondrial eytochrome oxidase submit （COI） gene region （450 bp） to investigate the genetic structure of the oriental river prawn （ Macrobrachium nipponense ） among nine populations from the Yangtze and Lancang Rivers. A total of 79 individuals were collected for this work. Eighty-nine nucleotides were found to be variable, resulting in 46 haplotypes. Among the nine populations, the population from Kunming shows the greatest level of variability （h = 1.000, π = 0.028）, whereas the population from Cbongqing exhibits the lowest level of variability （h = 0.700,π = 0.008）. Analysis of molecular variance suggested that of the total genetic diversity, 9.66% was attributable to inter-population diversity and the remainder （90.34%） to differences within populations. A molecular phylogenetic tree constructed using the Neighbor-joining （N J） method showed that the 46 haplotypes were assigned to two clades associated with geographic regions. These results provide basic information for the conservation and sustainable exploitation of this species.

Population Genetic Structure in Apricot (Prunus armeniaca L.) Cultivars Revealed by Fluorescent-AFLP Markers in Southern Xinjiang,China

Population-wide genetic structure was studied using fluorescent-AFLP markers on 85 apricot （Prunus armeniaca L.） cultivars collected from Kuche, Kashi, Hetian in the Tarim Basin, southern Xinjiang Uygur Autonomous Region of China. The purpose of this study was to determine the genetic structure and genotypic diversity among the different eco-geographical populations. Based on the results from this study, 8 pairs of fluorescent-AFLP primers showed clear electrophoregram and high polymorphism amongst the 64 pairs of EcoR Ⅰ/Mse Ⅰ （Mse Ⅰ - a FAM fluorescent marked primer） primers screened. There was a significant polymorphic difference for the same primer pair in different populations and for the same population with different primer pairs. The percentage of polymorphic loci （P） at species level was higher than Kuche, Hetian, Kashi population levels, respectively. The Nei＇s gene diversity index （H） and Shannon＇s information index （I） at species level were higher than those of Kuche, Hetian, and Kashi at population level, respectively. H and I of Kuche population were the highest amongst the three populations. Apricot population genetic diversity was found mainly within the population, Genetic differentiation coefficient between populations （GST） was 0.0882. Gene flow Nm between the populations was 5.1689. Population genetic identity was between 0.9772-0.9811 and genetic distance was between 0.0191-0.0232. These results further indicated that the similarity between populations was higher and the genetic distance between populations was smaller. The UPGMA cluster analysis indicates that the geographical populations at Kuche, Kashi, Hetian were relatively independent Mendelian populations. Concurrently, there was also partial gene exchange between the populations. All the evidences indicated that the genetic diversity in Kuche population was the highest, suggesting that it could be a transition population from wild apricot to cultivated apricot. There were abundant genetic diversities in apricot cultivar populations in southern Xinjiang, China, which provide promising germplasm for further breeding and theoretical basis for biodiversity conservation and utilization for apricot population in this area.

Phylogenetic Relationship of the Firefly,Diaphanes pectinealis(Insecta,Coleoptera,Lampyridae) Based on DNA Sequence and Gene Structure of Luciferase

Diaphanes is the fourth largest genus in Lampyridae, but no luciferase gene from this genus has been reported. In this paper, by PCR amplification of the genomic DNA, the luciferase gene of Diaphanes pectinealis, which is the first case from Diaphanes, was identified and sequenced. The luciferase gene from D. pectinealis spans 1958 base pairs （bp） from the start to the stop codon, including seven exons separated by six introns, and encoding a 547-residuelong polypeptide. Its deduced amino acid sequence showed high protein similarity to those of the Lampyrini tribe （93 - 94% ） and the Cratomorphini tribe （92%）, while low similarity was found with the North American firefly Photinus pyralis （83%） of the Photinini tribe within the same subfamily Lampyrinae. The phylogenetic analysis performed with the deduced amino acid sequences of the luciferase gene further confirms that D. pectinealis, Pyrocoelia, Lampyris, Cratomorphus, and Photinus belong to the same subfamily Lampyrinae, and Diaphanes is closely related to Pyrocoelia, Lampyris, and Cratomorphus. Furthemore, the phylogenetic analysis based on the nucleotide sequences of the luciferase gene indicates Diaphanes is a sister to Lampyris. The phylogenetic analyses are partly consistent with morphological （Branham ＆ Wenzel, 2003） and mitochondrial DNA analyses （Li et al, 2006）.

Genetic diversity and population structure of Marsh Grassbird (Locustella pryeri sinensis) in China

We used sequences of mitochondrial control region （807bp） in 75 samples from three breeding colonies and one wintering population to investigate the genetic diversity and population structure of Marsh Grassbird （Locustella pryeri sinensis） in different regions of China. Marsh Grassbird retained a moderate amount of haplotype （0.759 ± 0.056） and nucleotide diversity （0.002）. The results of FST among 3 phy-logeographic units and ФST between breeding and wintering sites revealed little evidence of genetic distinction between different colonies. Neither UPGMA tree structure analysis nor Network picture analysis showed obvious divergence between populations at different locations. Analysis of molecular variance also showed a lack of regional subdivision within Locustella pryeri sinesis, 98.5% of source of variation within populations and only 1.5% among populations. The neutrality test showed negative Fu’s FS value, which, in combination with detection of the mismatch distribution, suggested that population expansion occurred in the evolu-tionary history of this species. This hypothesis was further supported by Tajima’s D test and Fu’s test （D = -1.80, p = 0.02; Fs = -22.11, p = 0.001）, this expansion was estimated to occur about 28,700 years ago.

Structure of the Bovine ACAD8 Gene and the Association of Its Polymorphism with the Production Traits

Acyl-coenzyme A dehydrogenases （ACAD） are a family of nuclear-coded, mitochondrial flavoenzymes that catalyze the alpha, and beta-dehydrogenation of fatty acids. The eighth member of this family, ACAD8 catalyzes the valine catabolism. In this study, the bovine ACAD8 full-length mRNA and genomic DNA sequence were obtained and its gene structure was determined through alignment of the genomic DNA sequence to the mRNA sequence. The mRNA sequence consisted of a 1,251 bp open reading frame （ORF） flanked by a 37 bp 5＇-untranslated region （UTR） and a 444 bp 3＇-UTR; and its full-length genomic DNA sequence was 13,814 bp in length and included 11 exons and 10 introns. One A-G single nucleotide polymorphism （SNP） was revealed at nucleotide 13,408 （GenBank accession No. DQ435445） in the bovine ACAD8 gene by sequencing the polymerase chain reaction （PCR） products of 6 randomly selected individuals from the sample population. Different genotypes were determined by restriction fragment length polymorphism （RFLP）. The association analysis of this SNP in bovine ACAD8 with production traits in 178 unrelated steers from 5 breeds showed that it had a significant effect on the daily gain and the beef tenderness （P〈0.05）. Cattle with the G allele grew more rapidly and the beef they produced was more tender than those with the A allele. Thus, this SNP of the bovine ACAD8 gene can be used as an indicator to improve the growth rate and the beef tenderness.

Analysis of Genetic Variation and Population Structure of Starch Synthesis-related Genes in Indica Rice Cultivars

In this study, 34 molecular markers of starch synthesis-related genes were used to evaluate the genetic variation and population structure of 87 indica rice cultivars from different countries and regions. The results showed that a total of 80 alleles were amplified using 34 primer pairs, with an average of 2.5 alleles per locus. The allele number varied from 2 to 6 among various cultivars. Shannon＇s diversity index of molecular markers varied from 0.303 to 0.796, with an average of 0.539. Polymorphism information content （PIC） varied from 0.084 to 0.658, with an average of 0.295. The genetic similarity coefficients of 87 indica rice cultivars ranged from 0.265 to 0.990, indicating significant genetic differences of starch synthesis-related genes among different cultivars, but the variation frequency of alleles varied among different cultivars. Population structure analysis showed that these 87 indica rice cultivars were divided into three categories. Genetic differences were small within the same category but great among different categories. Moreover, indica rice cultivars with simple genetic components accounted for 39.1% and those with complex genetic background accounted for 60.9%. This study may not only provide theoretical basis for genetic improvement of rice starch quality, but also lay a solid foundation for subsequent association analysis of rice quality-related traits.

Morphological Structure and Genetic Mapping of New Leaf-Color Mutant Gene in Rice (Oryza sativa)

Leaf-color mutations are a widely-observed class of mutations, playing an important role in the study of chlorophyll biosynthesis and plant chloroplast structure, function, genetics and development. A naturally-occurring leaf-color rice mutant, Baihuaidao 7, was analyzed. Mutant plants typically exhibited a green-white-green leaf-color progression, but this phenotype was only expressed in the presence of a stress signal induced by mechanical scarification such as transplantation. Prior to the appearance of white ~eaves, mutant plant growth, leaf color, chlorophyll content, and chloroplast ultrastructure appeared to be identical to those of the wild type. After the changeover to white leaf color, an examination of the mutated leaves revealed a decrease in total chlorophyll, chlorophyll a, chlorophyll b, and carotenoid content, a reduction in the number of chloroplast grana lamella and grana, and a gradual degradation of the thylakoid lamellas. At maturity, the mutant plant was etiolated and dwarfed compared with wild-type plants. Genetic analysis indicated that the leaf mutant character is controlled by a recessive nuclear gene. Genetic mapping of the mutant gene was performed using an F2 population derived from a Baihuaidao 7 ~ Jiangxi 1587 cross. The mutant gene was mapped to rice chromosome 11, positioned between InDel markers L59.2-7 and L64.8-11, which are separated by approximately 740.5 kb. The mutant gene is believed to be a new leaf-color mutant gene in rice, and is tentatively designated as gwgl.

Effects and mechanism of adenovirus-mediated phosphatase and tension homologue deleted on chromosome ten gene on collagen deposition in rat liver fibrosis

AIM To evaluate the effects of phosphatase and tension homologue deleted on chromosome ten(PTEN) gene on collagen metabolism in hepatic fibrosis and the underlying mechanisms.METHODS rat primary hepatic stellate cells(HSCs) and human LX-2 cells were transfected with adenovirus containing c DNA constructs encoding wild-type PTEN(Ad-PTEN), PTEN mutant G129 E gene(Ad-G129E), and r NA interference constructs targeting the PTEN sequence PTEN short hairpin r NA to up-regulate and downregulate the expression of PTEN. HSCs were assayed using fluorescent microscopy, real-time polymerase chain reaction, and western blotting. Moreover, a CCl_4-induced rat hepatic fibrosis model was established to investigate the in vivo effects. Hematoxylin and eosin, and Masson's trichrome were used to assess the histological changes. The expression of collagen Ⅰ and Ⅲ was assessed using immunohistochemistry and western blot analysis.RESULTS Elevated expression of PTEN gene reduced serum levels of alanine transaminase and aspartate transaminase, decreased collagen deposition in the liver, and reduced hepatocyte necrosis. In contrast, knockdown of PTEN expression had an opposite effect, such as increased collagen deposition in the liver, and was molecularly characterized by the increased expression of matrix metalloproteinase(MMP)-13(P < 0.01) and MMP-2(P < 0.01), as well as decreased expression of the tissue inhibitor of metalloproteinase(TIMP)-1(P < 0.01) and TIMP-2(P < 0.01).CONCLUSION These data indicated that gene therapy using recombinant adenovirus encoding PTEN might be a novel way of treating hepatic fibrosis.

Spatial distribution characteristics of bacterial community structure and gene abundance in sediments of the Bohai Sea

This study investigated differences in the community structure and environmental responses of the bacterial community in sediments of the Bohai Sea.Illumina high-throughput sequencing technology and real-time PCR were used to assay the bacterial 16S rRNA genes in the surface sediments of 13 sampling stations in the Bohai Sea.The results showed that sediments at the majority of the 13 sampling stations were contaminated by heavy metal mercury.The main phyla of bacteria recorded included Proteobacteria(52.92%),Bacteroidetes(11.76%),Planctomycetes(7.39%),Acidobacteria(6.53%)and Chloroflexi(4.97%).The genus with the highest relative abundance was Desulfobulbus(4.99%),which was the dominant genus at most sampling stations,followed by Lutimonas and Halioglobus.The main factors influencing bacterial community structure were total organic carbon,followed by depth and total phosphorus.The content of lead,cadmium,chromium,copper and zinc had a consistent effect on community structure.Arsenic showed a negative correlation with bacterial community structure in most samples,while the impact of mercury on community structure was not significant.The bacterial community in sediment samples from the Bohai Sea was rich in diversity and displayed an increase in diversity from high to low latitudes.The data indicated that the Bohai Sea had abundant microbial resources and was rich in bacteria with the potential to metabolize many types of pollutants.

Cloning the Structure Genes and Expression the N Gene of Porcine Epidemic Diarrhea Virus DX

The structure genes spike(S) ,nucleocapsid(N) ,membrane(M) ,small membrane(sM) of a porcine epidemic diarrhea virus(PEDV) strain DX isolated in Gansu province,North-west of China,were cloned,sequenced and compared with published sequences of PEDV strains. The nucleotide sequences encoding the entire S,sM,M and N genes open reading frame(ORF) of DX were 4 152,231,681 and 1 326 bases long respectively. There were transcription regulatory sequences(TRSs) upstream of the initiator ATG of the S,N and M genes. The amino acids sequences of S,M and N contained 30,3 and 7 potential asparagine(N) -linked glycosylation sites. Homologous analysis and phylogenetic trees showed that DX had the closest relationship with strains LJB/06,JS-2004-2Z and CH/HLJH/06 that were also isolated from China and indicated the prevalence of some PEDV isolates in China were widespread since the JS-2004-2Z strain originated from the south of the China,and LJB/06 and CH/HLJH/06 were isolated from northeast China. The N gene was cloned using two primers which contained Nco I and BamH I restriction enzyme sites and subcloned into expression vector pET30a. The recombinant plasmid was then transformed into E.coli Rossta. SDS-PAGE showed there was a protein of about 55kDa as expected and Western blot indicated the N protein had biological activity.

The promoter analysis of the human C17orf25 gene, a novel chromosome 17pl3.3 gene

The human C17orf25 gene (Accession No. AF177342) is one of thirteen genes cloned from a region displaying a high score of loss of heterozygosity within chromosome 17pl3.3 in human hepatocellular carcinoma in China[l]. To unveil the underlying mechanisms for the transcription regulation of this gene and understand its implication to the hepatocellular carcinogenesis, we looked into the relevant aspects by both bioinformatic and experimental executions. We found: 1, The abundant expression of the C17orf25 gene was evident in all the cell lines and tissue samples tested, showing little hepatoma-selectivity; 2, Its transcription starts at a single site, locating at -60 from the translation initiation codon; 3, A 58 bp fragment containing the transcription start, extending from -112 to -55, represents the minimal promoter; 4, The consensus sequence within this fragment recognized by SP1 contributes predominantly to the activity of the minimal promoter; 5, The bioinformatic analysis suggests that the C17orf25 gene may encode a protein in the family of the glyoxalase. Our data has provided some deep insight into both function and regulation of the C1 7orf25 gene in the context of the normal liver and hepatocellular carcinoma.

Cloning and characterization of a novel gene (C17orf25) from the deletion region on chromosome 17p13.3 in hepatocelular carcinoma

Using a combination of hybridization of PAC to a cDNA library and RACE technique, we isolated a novel cDNA, designated as C17orf25 (Chromosome 17 open rea(ling frame 25, previously named it HC71A), from the deletion region on chromosome 17p13.3. The cDNA encodes a protein of 313 amino acids with a calculated molecular mass of 34.8 kDa. C17orf25 is divided into 10 exons and 9 introns, spanning 23 kb of genomic DNA. Northern blot analysis showed that the mRNA expression of C17orf25 was decreased in hepatocellular carcinoma samples as compared to adjacent noncancerous liver tissues from the same patients. The transfection of C17or25 into the hepatocellular carcinoma cell SMMC7721 and overexpression could inhibit the cell growth. The above results indicate that C17orf25 is a novel human gene, and the cloning and preliminary characterization of C17orf25 is a prerequisite for further functional analysis of this novel gene in human hepatocellular carcinoma.

Mapping of metastasis suppressor genes for prostate cancer by microcell-mediated chromosome transfer

Aim:To identify the metastasis suppressor genes for prostate cancer.Methods:A copy of human chromosomeswas introduced into the highly metastatic Dunning R-3327 rat prostate cancer cells by the use of microcell-mediatedchromosome transfer.Relationships between the size of human chromosomes introduced into microcell hybrid clonesand the number of lung metastases produced by the clones were analyzed to determine which part of human chromo-somes contained the metastasis suppressor gene(s)for prostate cancer.To determine portions of human chromosomesintroduced,G-banding chromosomal analysis,fluorescence in sim hybridization analysis,and polymerase chain reac-tion analysis were performed.Results:Each of microcell hybrid clones containing human chromosomes 7,8,10,11,12,or 17 showed decreased ability to metastasize to the lung without any loss of tumorigenicity.This demonstratesthat these human chromosomes contain metastasis suppressor genes for prostate cancer.Spontaneous deletion of portionsof human chromosomes was observed in the human chromosome 7,10,11,12,and 17 studies.In the human chromo-some 8 study,irradiated microcell-mediated chromosome transfer was performed to enrich chromosomal arm deletionsof human chromosome 8.Molecular and cytogenetic analyses of microcell hybrid clones demonstrated that metastasissuppressor genes on human chromosomes were located on 7q21-22,7q31.2-32,8p21-12,10q11-22,11p13-11.2,12p11-q13,12q24-ter,and 17pter-q23.KAII and MKK4/SEKI were identified as metastasis suppressor genes from11p11.2 and 17p12,respectively.Conclusion:This assay system is useful to identify metastasis suppressor gene(s)for prostate cancer.

Bioinformatics analysis of structure and function in the MRP gene family and its expression in response to various drugs in Bursaphelenchus xylophilus

Genes homologous to members of the MRP gene family in Caenorhabditis elegans are important in drug resistance.To further explore the molecular mechanism of drug resistance in pine wood nematode(Bursaphelenchus xylophilus),we used bioinformatics approaches to analyze genomic data for B.xylophilus and identified Bx-MRP genes.We predicted the structure and function of the genes and encoded proteins.Using bioinformatics programs to predict and analyze various properties of the predicted proteins,including hydrophobicity,transmembrane regions,phosphorylation sites,and topologically isomeric structures,of these Bx-MRP genes,we determined that they function in transmembrane transport.From the results of RT-qPCR,the Bx-MRP family members confer significant differential resistance to different drug treatments.After treatment with different concentrations of emamectin benzoate,avermectin and matrine,the expression of each gene increased with increasing drug concentrations,indicating that the family members play a positive role in the regulation of multidrug resistance.

Genetic Variability and Population Structure of Ark Shell in Japan

Ark shell Scapharca kagoshimensis is one of the commercially important bivalve resources in East Asia. In Japan, the mass production method for its natural seedlings was developed in the 1880s, and they had been transplanted to an array of the major fishing areas. It has been therefore concerned with its genetic disturbance among not only current but also former fishing areas in Japan. This study was undertaken to ascertain its genetic diversity and population structure in East Asia by means of nucleotide sequence analysis of a 555-bp portion of the mitochondrial DNA COI gene. Of 225 individuals collected from 8 populations and 1 population in Japan and Korea, respectively, a total of 59 haplotypes, including 14 common haplotypes, were found, and Japan and Korea shared 3 common haplotypes. In Japan, the haplotype diversity and nucleotide diversity ranged from 0.65 to 0.93 and from 0.22% to 0.59%, respectively, reflecting relatively high levels of genetic diversity. The values in Korea were determined to be 0.45% and 0.19%, respectively, indicating significantly lower genetic diversity compared with that in Japan. Mismatch distribution analysis and neutrality tests showed a recent history of multiple types of reproduction and signals of demographic change in each population. These results suggest that S. kagoshimensis has experienced rapid population growth or reduction in population size such as a bottleneck in a short period.

Dissemination and Genetic Structure of Carbapenemase Encoding Genes (bla_OXA-23and bla_OXA-24) in <i>Acinetobacter baumannii</i>from Southern Texas

Acinetobacter baumannii is one of the most important human pathogens causing a variety of nosocomial infections. Carbapenem antibiotics have been primarily used to treat the A. baumannii infections. However, carbapenem resistant A. baumannii producing carbapenemases causes serious treatment problems worldwide. Outbreaks of carbapenem resistant isolates have reported in some area of the United States, but their dissemination and genetic structure of the carbapenemase encoding genes are currently little known. To understand outbreaks, dissemination, and genetic structure of the carbapenemase encoding genes in Southern Texas, 32 clinical isolates collected from Austin and Houston, TX were characterized. Twenty-eight of 32 isolates were resistant to all tested β-lactam antibiotics including carbapenem (imipenem and meropenem). Three of them carried blaOXA-23 as a part of Tn2008 integrated into a known plasmid (pACICU2) and all others carried blaOXA-24 flanked by XerC/XerD-like recombinase binding sites that were adjoined by DNA sequences originated from multiple plasmids. Genotype analysis revealed that the 25 isolates carrying blaOXA-24 were all identical genotypes same as a representative isolate carrying blaOXA-24 from Chicago, IL but the 3 isolates carrying blaOXA-23 was a distinct genotype as compared with isolates carrying blaOXA-23 from Chicago, IL and Washington, D.C. Each of the blaOXA-23 and blaOXA-24 was transferred to carbapenem susceptible A. baumannii and E. coli with similar minimal inhibitory concentration (MIC) of carbapenem as that of their parental isolates but significantly lower levels of MIC in E. coli. Overall results suggest that a unique strain carrying blaOXA-23 and a similar strain carrying blaOXA-24 as seen in other geographic areas are currently disseminated in Southern Texas.

Genomic structure analysis of SNC6, a progesterone-receptor associated protein gene, and cloning and characterization of its 5＇-flanking region

Objective: To analyze the genomic structure of SNC6, a progesterone\|receptor associated protein gene and its regulatory elements in its 5'\|flanking region. Methods: Genomic sequence from GenBank database (accession number: Z98048) covering the whole SNC6 gene was used to analyze the genomic structure of SNC6 and design primers for PCR amplification of its 5'\|flanking region. A 1894 bp fragment of the 5'\|flanking region \{(-1814\} to +75) was cloned by PCR using genomic DNA from a healthy donor peripheral blood lymphocyte as template. This fragment, as well as 3 shorter derivative fragments (1423 bp, 632 bp and 416 bp, which correspond to -1344 to +75, -552 to +75 and -337 to +75 respectively), were subcloned into pGL2 series luciferase reporter vectors. These constructs were introduced into colorectal cancer cell line SW620 for transient expression of reporter gene and luciferase activities were measured. Results: The genomic structure analysis showed there are 12 exons for SNC6 gene, which spans 32017 bp (nt71529 to nt39513 in Z98048 sequence). All transfected SW620 cells with the above 5\|flanking region\|containing constructs showed luciferase activities. The highest luciferase activities were measured in transfected cells with vectors containing 1894 bp fragments, and the lowest luciferase activities were measured in transfected cells with vectors containing 416 bp fragments. Luciferase activities were higher in transfected cells with vectors containing 632 bp fragments than that in transfected cells with vectors containing 1423 bp fragments. Conclusion: The basic transcription\|promoting element (promoter) for SNC6 expression resides between 0 to -337, and two transcription\|enhancing elements (enhancer) resides between -337 to -552 and -1344 to -1814, whereas one transcription\|inhibiting element (silencer) exists between -552 to -1344.

The heterodimeric structure of heterogeneous nuclear ribonucleoprotein C1/C2 dictates 1,25-dihydroxyvitamin D-directed transcriptional events in osteoblasts

Heterogeneous nuclear ribonucleoprotein （hnRNP） C plays a key role in RNA processing but also exerts a dominant negative effect on responses to 1,25-dihydroxyvitamin D （1,25（OH）2D） by functioning as a vitamin D response element-binding protein （VDRE-BP）. hnRNPC acts a tetramer of hnRNPC1 （huC1） and hnRNPC2 （huC2）, and organization of these subunits is critical to in vivo nucleic acid-binding. Overexpression of either huC1 or huC2 in human osteoblasts is sufficient to confer VDRE-BP suppression of 1,25（OH）2D-mediated transcription. However, huC1 or huC2 alone did not suppress 1,25（OH）2D-induced transcription in mouse osteoblastic cells. By contrast, overexpression of huC1 and huC2 in combination or transfection with a bone-specific polycistronic vector using a ＂self-cleaving＂ 2A peptide to co-express huC1/C2 suppressed 1,25D-mediated induction of osteoblast target gene expression. Structural diversity of hnRNPC between human/NWPs and mouse/rat/rabbit/dog was investigated by analysis of sequence variations within the hnRNP CLZ domain. The predicted loss of distal helical function in hnRNPC from lower species provides an explanation for the altered interaction between huC1/C2 and their mouse counterparts. These data provide new evidence of a role for hnRNPC1/C2 in 1,25（OH）2D-driven gene expression, and further suggest that species-specific tetramerization is a crucial determinant of its actions as a regulator of VDR-directed transactivation.

CHROMOSOME 17P MAY HARBOR MULTIPLE TUMOR SUPPRESSOR GENES ASSOCIATED WITH PRIMARY GLIOBLASTOMA MULTIFORME

Objective: To investigate whether deletion of chromosome 17 is involved in the carcinogenesis of primary glioblastoma multiforme and to localize the possible common deletion region in the aforementioned chromosome. Methods: Polymerase chain reaction-based microsatellite analysis was used to assess loss of heterozygosity (LOH) on chromosome 17 in 20 primary glioblastoma multiforme (GBM). Fifteen fluorescent dye-labeled polymorphic markers were used. Results: Thirteen of twenty (65%) GBM displayed LOH on at least one marker of chromosome 17p. Two tumors showed either LOH or non-informativeness on all markers tested. The most frequent LOH was observed at loci including D17s799 (53.3%), D17s1852 (53.8%), D17s938 (63.20/o), D17s831 (55.6%). The loci D17s831 (on 17p13) and D17s799–D17s1852 (17p11.2–p12) are distal and proximal to p53 respectively. The frequencies of LOH at all loci examined on chromosome 17q were relatively low (<30%). None of informative loci exhibited microsatellite instability in this study. Conclusion: Loss of genetic material on chromosome 17p may play an important role in the pathogenesis of GBM. Besides the well-known TSG p53 on 17p, other unknown TSCs associated with GBM may be present on the chromosomal regions 17p13 and 17p11.2–p12, which are distal and proximal to p53 respectively.