Association of Genetic Variation in the Promoter Region of OXTR with Differences in Social Affective Neural Processing

Evidence supports the involvement of oxytocin in social behavior. The oxytocin receptor gene (OXTR) has been associated with differences in social brain function and risk for autism. Motivated by recent work, we investigated the effect of variation in the common functional rs2268498 T/C polymorphism in the promoter region of OXTR on neural responses to fear expressions. 46 healthy subjects were divided into genotype groups of C carriers (n = 32) and TT ho-mozygous (n = 14) and neural activity was measured during the recognition of fear and neutral expressions. Results showed that during the recognition of fear expressions, the TT genotype group exhibited increased responding in the inferior occipital gyrus, considered important for face processing, compared to carriers of the C allele (P < 0.005;cluster corrected for whole brain), an effect not found for neutral faces. These results indicate the impact of this OXTR genetic variant on individual differences in social affective neural processing.

Shallow Genetic Structure of Pholis fangi in Bohai Sea and Yellow Sea Inferred from mt DNA Control Region

Pholis fangi is an ecologically important fish species inhabiting Chinese coastal waters of Yellow Sea and Bohai Sea. To investigate the genetic diversity and population genetic structure of P. fangi, a fragment of 487 bp in the first hypervariable region (HVR-1) of the mitochondrial DNA (mtDNA) control region was sequenced for 181 individuals collected from Bohai Sea and Yel- low Sea. A total of 18 polymorphic sites were detected, which defined 25 haplotypes. A moderate level of haplotype diversity (h = 0.7052) and a low level of nucleotide diversity (π= 0.0028) were detected. Both the phylogenetic tree and the haplotype network showed no significant genealogical structure difference among sampling locations. Pairwise FST comparison and hierarchical mo- lecular variance analysis (AMOVA) revealed that no significant genetic structure difference existed throughout the investigated re- gions, suggesting a high gene exchange among different populations. The results of neutrality test and mismatch distribution analysis indicated that a late Pleistocene population expansion (38000 127000 years ago) happened. Seasonal schooling migration may con- tribute to the genetically homogeneous population structure of the species.

A Method for Upscaling Genetic Parameters of CERES-Rice in Regional Applications

To upscale the genetic parameters of CERES-Rice in regional applications, Jiangsu Province, the second largest rice producing province in China, was taken as an example. The province was divided into four rice regions with different rice variety types, and five to six sites in each region were selected. Then the eight genetic parameters of CERES-Rice, particularly the four parameters related to the yield, were modified and validated using the Trial and Error Method and the local statistical data of rice yield at a county level from 2001 to 2004, combined with the regional experiments of rice varieties in the province as well as the local meteorological and soil data （Method 1）. The simulated results of Method 1 were compared with those of other three traditional methods upscaling the genetic parameters, i.e., using one-site experimental data from a local representative rice variety （Method 2）, using local long-term rice yield data at a county level after deducting the trend yield due to progress of science and technology （Method 3）, and using rice yield data at a super scale, such as provincial, ecological zone, country or continent levels （Method 4）. The results showed that the best fitness was obtained by using the Method 1. The coefficients of correlation between the simulated yield and the statistical yield in the Method 1 were significant at 0.05 or 0.01 levels and the root mean squared error （RMSE） values were less than 9% for all the four rice regions. The method for upscaling the genetic parameters of CERES-Rice presented is not only valuable for the impact studies of climate change, but also favorable to provide a methodology for reference in crop model applications to the other regional studies.

Studies on Genetic Transformation of Fresh-cut Chrysanthemum Using DREB1A Promoted by Stress-induced Promoter rd29A

In this study, DERB1A transcription factor and stress-induced promoter rd29A were isolated respectively and amplified from Arabidopsis thaliana, se- quenced and analyzed by DNAsis. In addition, the stress-induced promoter rd29A was utilized to construct the plant expression vector of DERB1A, which was transformed into Agrobacterium tumefaciens. Furthermore, the transgenic regeneration system of fresh-cut chrysanthemum from callus to plantlets was established successfully. On this basis, chrysanthemum leaf-disc explants were genetically transformed with Agrobacterium-mediated method. Two positive transgenie plantlets were obtained in vitro. Based on PCR detection, DREB1A transcription factor was integrated into chrysanthemum genome, which laid the foundation for breeding new transgenie cultivars of fresh-cut chrysanthemum with high comprehensive stress resistance, good cmalitv and high field.

Analysis of SNPs in the Promoter Region of the Growth Hormone(GH) Gene in Mini-pigs

Single nucleotide polymorphisms(SNPs) of the growth hormone(GH) gene were investigated in six pig breeds,consisting of four mini-pig breeds(Wuzhishan,Bama,Xiang and Tibet pig),and two others(Dahlan and Landrace pig).Three pairs of primers for promoter regions of the GH gene were designed on the basis of the pig genomic sequence and SNPs were detected by the PCR-SSCP method.The results indicated three mutations in the 5’-flanking region.The analysis results showed that the frequencies of allele A and D in four mini-pig breeds were higher than that in other breeds at a locus within the 5’-flanking region(P【0.05).These results suggest that differences in body size may be associated with these SNPs of 5’-flanking region and amino acid mutation of the signal peptide of GH in these pig breeds.

Cloning and Analysis of the Promoter Region of Rat uPA Gene

Objective To clone and analyze the promoter sequence of rat urokinase plasminogen activator protein gene. Methods The genomic DNA was extracted from rat testicular tissue. According to urokinase plasminogen activator, the gene sense primer and antisense primer of uPA gene were designed and synthesized, then Touch-Down PCR were performed. After proper purification, the PCR product was sequenced, analyzed with the promoter prediction software and compared with the DNA sequence of rattuas urokinase plasminogen activator. Results The cloned uPA gene was about 1 572 bp in length, which contained a full open-reading frame with 21 bp in length exons, and the upper region of transcriptional start was 1 551 bp in length which was eucaryon transcriptional control area. The 5＇ UTR had a promoter region including a non-responsive TATA-box. Not only the GC-box binding region was found in this gene, but also active protein I （AP1） and SP1 were seen in other regions. Conclusion A 1 572 bp uPA gene fragment （GenBank accession No.X65651） was obtained from rat genomic DNA library, containing eucaryon transcriptional control area with a promoter region, non-conspicuous TATA-box, GC-box and an extron. A non-responsive TATA-box is located at the upper -30 region.

Population Genetic Analysis of Sillago nigrofasciata (Perciformes:Sillaginidae) Along the Coast of China by Sequencing Mitochondrial DNA Control Region

Sillago nigrofasciata, a small to moderate size nearshore species, is newly found along the eastern and southern coasts of China. The present study is carried out in order to analyze the population genetics of the S. nigrofasciata. The control region sequence of mitochondrial DNA revealing 73 haplotypes were obtained from 162 individuals collected at 8 locations along the coast of China. The whole S. nigrofasciata population along the coast of China showed a low nucleotide diversity(0.012) and a high population diversity(haplotype diversity)(0.943), and all the 8 local populations showed low nucleotide diversities(0.014 – 0.001), suggesting the protective measures are effective. The haplotypes of the 8 local populations were widely distributed in haplotype network diagram and neighbor-joining phylogenetic tree, while no branch associating with sampling locations was detected. Recent gene flow analysis showed asymmetric gene exchanges among local populations. The pairwise FST values and unweighted pair-group method with arithmetic mean(UPGMA) tree revealed a certain amount of genetic difference among local populations. Moreover, analysis of molecular variance(AMOVA) reflected genetic differences between hypothetical subdivision groups. Neutral test and mismatch distribution of pairwise nucleotide suggested S. nigrofasciata may have experienced recent population expansion events. The historical geographic events associating with ice age may be the main explanation to the heterogeneity among local populations with short geographic distances, and the homogeneity among local populations with long geographic distances.

Evaluation of Uncertainty in Measuring the Content of CaM V35S Promoter in Genetically Modified Soybean,GTS40-3-2,by Real-time Quantitative PCR

[ Objective ] This study aimed to investigate the major contributors to the measurement uncertainty in quantitative analysis of genetically modified ingreclients and improve the quality of quantitative detection of genetically modified components. [ Method] The content of CaMV35S promoter （parameter） in GTS40- 3-2 soybean powder samples was measured to estimate the measurement uncertainty preliminarily. [ Result] Type A uncertainty （uA） ＇ type B uncertainty （uB） and combined standard uncertainty （Uc） were 0.0 004, 0.002 and 0.002, respectively. At a confidence level ofp = 95% and freedom degree of Voff = 3 251, coverage factor k = 1.96, expanded uncertainty U = 0.004. The final measurement result was C = 0.028 ± 0. 004, which was dose to the conventional true value （0.03）. Thus, the measurement uncertainty was relatively small, indicating a high quality of measurement. In this study, uncertainty evaluation indicated that the deviation of micro liquid transfer made the greatest contribution to the measurement uncertainty. [ Cludusion ] The deviation of micro liquid transfer should be reduced to im- prove the quality of measurement.

The Genetic Structure and Diversity of Repomucenus curvicornis Inhabiting Liaoning Coast Based on Mitochondrial COⅠ Gene and Control Region

[Object] This study was conducted to explore the genetic diversity and structure of the wild Repomucenus curvicornis inhabiting Liaoning Coast, China. [Method] The mitochondrial COⅠ gene and control region（CR） were PCR amplified from the wild R. curvicornis populations from the Liaodong Bay（n=22） and the northern Yellow Sea（n=18）, sequenced and analyzed for genetic diversity. [Result] The contents of A, T, C and G of 624 bp COⅠ gene were 24.09%, 31.04%, 25.28%, and 19.59%, and those of 460 bp CR fragment were 32.96%, 32.80%, 14.86% and 19.38%, respectively. The total number of variable sites, average number of nucleotide differences（ k）, haplotype diversity（H） and nucleotide diversity（π） based on COⅠ gene were 38, 4.67,（0.96±0.02） and（0.007 5±0.004 2）, and those based on CR fragment were 26, 3.35,（0.97 ±0.02） and（0.007 3±0.004 3）, respectively. Based on mitochondrial COⅠ gene and CR, the genetic diversity of Liaodong Bay population was lower than that of the northern Yellow Sea population. The AMOVA analysis based on CR fragments revealed almost significant genetic divergence between the Liaodong Bay and the northern Yellow Sea populations, while there was no significant genetic divergence based on COⅠ gene. The results showed that CR and COⅠ gene are effective molecular markers for detecting the genetic diversity of R. curvicornis population, while CR is more reliable than COⅠ gene in detecting the genetic structure. [Conclusion] CR is an appropriate marker for genetic analysis of marine fish population.

Genetic map construction and functional characterization of genes within the segregation distortion regions(SDRs)in the F_(2:3) populations derived from wild cotton species of the D genome

Background:Segregation distortion(SD)is a common phenomenon among stable or segregating populations,and the principle behind it still puzzles many researchers.The F2:3 progenies developed from the wild cotton species of the D genomes were used to investigate the possible plant transcription factors within the segregation distortion regions(SDRs).A consensus map was developed between two maps from the four D genomes,map A derived from F2:3 progenies of Gossypium klotzschianum and G.davidsonii while Map B from G.thurberi and G.trilobum F2:3 generations.In each map,188 individual plants were used.Results:The consensus linkage map had 1492 markers across the 13 linkage groups with a map size of 1467.445 cM and an average marker distance of 1.0370 cM.Chromosome D502 had the highest percentage of SD with 58.6%,followed by Chromosome D507 with 47.9%.Six thousand and thirty-eight genes were mined within the SDRs on chromosome D502 and D507 of the consensus map.Within chromosome D502 and D507,2308 and 3730 genes were mined,respectively,and were found to belong to 1117 gourp out of which 622 groups were common across the two chromosomes.Moreover,genes within the top 9 groups related to plant resistance genes(R genes),whereas 188 genes encoding protein kinase domain(PF00069)comprised the largest group.Further analysis of the dominant gene group revealed that 287 miRNAs were found to target various genes,such as the gra-miR398,gramiR5207,miR164a,miR164b,miR164c among others,which have been found to target top-ranked stress-responsive transcription factors such as NAC genes.Moreover,some of the stress-responsive cis-regulatory elements were also detected.Furthermore,RNA profiling of the genes from the dominant family showed that higher numbers of genes were highly upregulated under salt and osmotic stress conditions,and also they were highly expressed at different stages of fiber development.Conclusion:The results indicated the critical role of the SDRs in the evolution of the key regulatory genes in plants.

Mutations in pre-core and basic core promoter regions of hepatitis B virus in chronic hepatitis B patients

AIM: To investigate the frequency of mutations in precore(pre-C) and basic core promoter(BCP) regions of hepatitis B virus(HBV) from Shanxi Province, and the association between mutations and disease related indexes.METHODS: One hundred chronic hepatitis B patients treated at Shanxi Province Hospital of Traditional Chinese Medicine were included in this study. PCRreverse dot blot hybridization and mismatch amplification mutation assay(MAMA)-PCR were used to detect the mutations in the HBV pre-C and BCP regions. HBV DNA content and liver function were compared between patients with mutant HBV pre-C and BCP loci and those with wild-type loci. The consistency between PCR-reverse dot blot hybridization and MAMA-PCR for detecting mutations in the HBV pre-C and BCP regions was assessed.RESULTS: Of the 100 serum samples detected, 9.38% had single mutations in the pre-C region, 29.17% had single mutations in the BCP region, 41.67% had mutations in both BCP and pre-C regions, and 19.79% had wild-type loci. The rates of BCP and pre-C mutations were 65.7% and 34.3%, respectively, in hepatitis B e antigen(HBe Ag) positive patients, and 84.6% and 96.2%, respectively, in HBe Ag negative patients. The rate of pre-C mutations was significantly higher in HBe Ag negative patients than in HBe Ag positive patients(χ~2 = 26.62, P = 0.00), but there was no significant difference in the distribution of mutations in the BCP region between HBeAg positive and negative patients(χ~2 = 2.43, P = 0.12). The presence of mutations in the pre-C(Wilcoxon W = 1802.5, P = 0.00) and BCP regions(Wilcoxon W = 2906.5, P = 0.00) was more common in patients with low HBV DNA content. Both AST and GGT were significantly higher in patients with mutant pre-C and BCP loci than in those with wild-type loci(P < 0.05). PCR-reverse dot blot hybridization and MAMA-PCR for detection of mutations in the BCP and pre-C regions had good consistency, and the Kappa values obtained were 0.91 and 0.58, respectively.CONCLUSION: HBe Ag negative patients tend to have HBV pre-C mutations. However, these mutations do not cause increased DNA copies, but associate with damage of liver function.

Genetic Analysis of the P1 Region of Human Enterovirus 71 Strains and Expression of the 55 F StrainVP1 Protein

Enterovirus 71 (EV71) is a member of the Entero-virus genus of the Picornaviridae family and is the major cause of Hand, foot, and mouth disease (HFMD) in children. Different strains from Gansu were cloned and the P1 protein was sequenced and analysed. Results indicate that there are three kinds of EV71 infections prevalent in Gansu. The VP1 protein from one of these strains, 55F, was expressed. The recombinant protein was expressed with high level and reacted specifically with the EV71 patient antibody, the recombinant protein was also applied to raise antiserum in rabbits and after the fourth injection a high titer of antiserum was detected by ELISA assay. These data are useful for further clarification of prevalent EV71 strains in the north of China at the molecular level and provide a basis for EV71 diagnosis.

Satellite Constellation Design with Multi-Objective Genetic Algorithm for Regional Terrestrial Satellite Network

Constellations design for regional terrestrial-satellite network can strengthen the coverage for incomplete terrestrial cellular network. In this paper, a regional satellite constellation design scheme with multiple feature points and multiple optimization indicators is proposed by comprehensively considering multi-objective optimization and genetic algorithm, and "the Belt and Road" model is presented in the way of dividing over 70 nations into three regular target areas. Following this, we formulate the optimization model and devise a multi-objective genetic algorithm suited for the regional area with the coverage rate under simulating, computing and determining. Meanwhile, the total number of satellites in the constellation is reduced by calculating the ratio of actual coverage of a single-orbit constellation and the area of targets. Moreover, the constellations' performances of the proposed scheme are investigated with the connection of C++ and Satellite Tool Kit(STK). Simulation results show that the designed satellite constellations can achieve a good coverage of the target areas.

Genetic variation of the small yellow croaker(Larimichthys polyactis)inferred from mitochondrial DNA provides novel insight into the fluctuation of resources

The small yellow croaker(Larimichthys polyactis)belongs to the family Sciaenidae,which is an offshore warm fish species and widely distributed in the western Pacific.In this study,the variation of genetic diversity and genetic differentiation among L.polyactis populations was analyzed by mitochondrial DNA control region.A total of 110 polymorphic sites were checked,which defined 134 haplotypes.High level of haplotype diversity(h=0.993±0.002)was detected in the examined range.Population genetic structure analyse(analysis of molecular variance,Fst)showed there were high gene flow among L.polyactis populations.The result showed that there were relatively high genetic diversity and low genetic differentiation among the Yellow Sea and the East China Sea populations,which can be attributed to diverse habitats,wide distribution range and high mutation rate of control region.Using phylogenetic methods,coalescent analyses(neutrality tests,mismatch distribution analysis,Bayesian skyline analyses)and molecular dating interpreted in conjunction with paleoclimatic and physiographic evidence,we inferred that the genetic make-up of extant populations of L.polyactis was shaped by Pleistocene environmental impacts on the historical demography of this species.Besides,relatively constant genetic diversity and larger effective population size were detected in recent L.polyactis population.The result showed that the fishing policy certainly,such as the summer closed fishing,played a role in protecting resources of L.polyactis.This study can offer a wealth of biological novelties which indicates genetic structure of L.polyactis population and provides the foundation for resources protection and policy setting.

A preliminary investigation on genetic diversity of Sousa chinensis in the Pearl River Estuary and Xiamen of Chinese waters

In this study, the mitochondrial DNA （mtDNA） control region and the mitochondrial cytochrome b gene of stranded Indo-Pacific humpback dolphin （Sousa chinensis） samples from the Pearl River Estuary and Xiamen waters were sequenced and analyzed. The result of mtDNA control region revealed 34 variable sites and four unique haplotypes （named as A, B, C and D） identified among the total samples from these two water areas, and the most common haplotype （A） was shared by 75% of the dolphins sampled from the two water areas. The haplotypic diversity （h） was 0.455 and the nucleodde diversity （π） was 0.0088. The phylogenetic analysis showed that the haplotype A, C, and D were closely related, but the haplotype B （unique for XM01 from Xiamen） was far from the other three. By scanning cytochrome b fragments, two haplotypes （A and B） were identified in these two water areas, and the most common haplotype （A） was shared by 91.67% individuals, while XM01 from Xiamen as the only exception. The data suggest that there is a possibility of gene exchange between the two populations in the Pearl River Estuary and Xiamen waters, and there possibly exists a unique maternal lineage in Xiamen waters.

Genetic analysis of selected Sargassum fusiforme (Harvey) Setchell (Sargassaceae, Phaeophyta) strains with RAPD and ISSR markers

Since the 1980s,Sargassum fusiforme has been cultivated in Zhejiang,South China,and nowadays it becomes one of the important commercial seaweeds in China.With traditions of eating habits in the East Asian countries,this brown alga is used as food,because it contained functional oligo/polysaccharides and chemical components,and was regarded playing roles in antioxidant activities and regulating immunology.Through over 15 years’selection,breeding and cultivation,we obtained three strains with good traits and testified their characters during the production,which included the cultivars with high yield and other two good characters,either all the selected strains were applied in the Sargassum production.To avoid confusion during the selection and nursery,it was preferred to establish one fingerprint for distinguishing the Sargassum cultivars from different strains.Random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR)methods were adopted to analyze the genetic diversities of the selected S.fusiforme strains.With that,one fingerprint with RAPD markers was constructed,and one sequence characterized amplifi ed region(SCAR)marker to S.fusiforme was obtained.It is indicated that the applied fingerprint could be valid in S.fusiforme genetic and germplasm justification,and will be positive to molecular marker assistance in its selection and cultivation.

Genetic Diversity and Species Identification of Cultivar Species in Subtribe Cucumerinae (Cucurbitaceae) Using RAPD and SCAR Markers

DNA polymorphism in the cultivar species;Cucumis sativus L., C. melo L. and Benincasa hispida Cogn. of subtribe Cucumerinae (Cucurbitaceae) in the four northeastern provinces of Thailand was examined by using RAPD technique. Twenty 10-mer primers were produced 212 RAPD fragments, ranging from approximately 120 to 2531 bp. The genetic similarities were estimated from banding profiles using a NTSYS* version 2.1 as a basis for dendrogram construction via the UPGMA method. Cluster analysis divided the taxa under study into 2 clades. Moreover, a RAPD marker: Cm (OPJ11700) was specified to C. melo, and this marker was converted into sequence characterized amplified region (SCAR) marker: Cm (SCJ11516). A pair of sequence-specific primer of clones Cm (OPJ11700) amplified a distinct single band of the same size as the RAPD clones. The SCAR marker was developed successfully to identify C. melo genotype.

Inter-and Intra-specific Genetic Diversity in Diospyros Using SCoT and IRAP Markers

The genus Diospyros contains more than 500 species, among them persimmon(Diospyros kaki Thunb.) is the most economically important species. However, the inter-and intra-specific genetic diversity remains unknown due to historic cultivation, domestication and introduction in different regions of China. In this study, we investigated the genetic diversity and relationship among 268 Diospyros accessions from different regions in China using start codon-targeted(SCo T) and inter-retrotransposon-amplified polymorphism(IRAP) markers. Among 268 samples,232 accessions were D. kaki varieties and 36 were closely related species, that is, D. lotus(21), D. glaucifolia(1), D. virginiana(5), D. kaki Thunb.var. silvestris(3), D. oleifera(5) and D. deyangnsis(1). In our study, a total of 90 and 97 polymorphic alleles were obtained from nine SCo T and nine IRAP markers, respectively, and these markers displayed rich polymorphisms for evaluation of the genetic diversity among 268 samples. A study on the relationship, genetic structure and analysis of molecular variance(AMOVA) showed that inter-and intra-specific gene exchanges were frequent among/within the samples from five regions. Moreover, the genetic distance between the Southwest China(SWC) and the other regions was relatively far;the samples from North China(NC), East China(EC), South and Central China(SCC), Northwest China(NWC) regions had a relatively close relationship. The unweighted pair group method with arithmetic means(UPGMA) and population structure analysis indicated that persimmons within geographic regions were clustered together, which revealed that the samples from various regions exhibited gene exchange, and the geographical proximity of persimmons resulted in a more frequent exchange. In addition, the persimmon had a close relationship with D. kaki var. silvestris, D. deyangnsis and D. oleifera species, and exhibited a far distance with respect to D. virginiana. These SCo T and IRAP markers will be helpful for understanding of the inter-and intra-specific genetic diversity of Diospyros germplasms, which play an important role in the germplasm efficient conservation, identification and utilization for Diospyros resources.

Advances in the Identification of Genetically Modified Rice with Real-time PCR and Multiplex PCR

In recent years, food security and safety have attracted increasing attention due to the worldwide research and development of genetically modified （GM） rice, and the controversy over the commercialization of GM rice. And the identification of GM rice is of great significance. Therefore, in the present study, the po- tential problems in the identification of GM rice with PCR were analyzed both at a technical level and from a theoretical perspective. In addition, PCR detection on the transgenic elements： promoter, terminator, internal reference gene and target gene was discussed, respectively. The possible solutions were proposed based on the principles of plant virology and genetic engineering.

Pre-stack seismic waveform inversion based on adaptive genetic algorithm

Pre-stack waveform inversion, by inverting seismic information, can estimate subsurface elastic properties for reservoir characterization, thus effectively guiding exploration. In recent years, nonlinear inversion methods, such as standard genetic algorithm, have been extensively adopted in seismic inversion due to its simplicity, versatility, and robustness. However, standard genetic algorithms have some shortcomings, such as slow convergence rate and easiness to fall into local optimum. In order to overcome these problems, the authors present a new adaptive genetic algorithm for seismic inversion, in which the selection adopts regional equilibrium and elite retention strategies are adopted, and adaptive operators are used in the crossover and mutation to implement local search. After applying this method to pre-stack seismic data, it is found that higher quality inversion results can be achieved within reasonable running time.