Isolation and Analysis of Rubber Hevein Gene and Its Promoter Sequence

Hevein, a lectin_like protein, is a major factor of lutoids in the latex of rubber trees ( Hevea brasiliensis Muell._Arg.). This factor is involved in coagulation of the latex and has the ability to bind chitin. The hevein gene with a length of 680 bp was cloned by the method of RT_PCR. Its promoter region with 1 306 bp of this gene was also isolated by genome walking, and its sequence included the typical TATA and CAAT boxes as well as the homologous sequence of abscisic acid (ABA) response elements. Expression of the hevein gene in the latex and leaves was detected by Northern blot. After treatment of the trees with ethylene and ABA, the results showed that the hevein gene was expressed principally in latex, and the expression could be induced by ethylene and ABA.

PROGRESS OF GREY SYSTEM MODELS

The progress of grey system models is reviewed, and the general grey numbers, the grey sequence op- erators and several most commonly used grey system models are introduced, such as the absolute degree of grey incidence model, the grey cluster model based on endpoint triangular whitenization functions, the grey cluster model based on center-point triangular whitenization functions, the grey prediction model of the model GM （ 1,1）, and the weighted multi-attribute grey target decision model.

Micro-heterogeneity of Storage Glutelin in Rice Seed

The storage glutelin in rice ( Oryza sativa L.) seeds could be separated at least into more than 13 acidic and 19 basic polypeptides by two dimensional electrophoresis. Rice glutelin might be mainly controlled by about six major genes according to the expression of glutelin polypeptides. The acidic polypeptide of glutelin could be clearly separated into two groups by peptide mapping and N-terminal amino acid sequence analysis. These two groups were just in accord with the GluA and GluB subfamilies exactly.

DISTRIBUTIONS OF TRIPLET CODONS IN MESSENGER RNA SECONDARY STRUCTURES

Analysis of the secondary structures of mRNAs which encode mature peptides shows that the location of each codon in mRNA secondary structure has a trend, which appears to be in agreement with the conformational property of the corresponding amino acid to some extent. Most of the codons that encode hydrophobic amino acids are located in stable stem regions of mRNA secondary structures, and vice versa, most of the codons that encode hydrophilic amino acids are located in flexible loop regions. This result supports the recent conclusion that there may be the information transfer between the three dimensional structures of mRNA and the encoded protein.

Segmental Duplications Are Common in Rice Genome

Segmental duplications on rice (Oryza sativa L.) chromosomes 8, 9, 11, and 12 were studied by examining the distributions of sequences resolved by 13 probes detecting multiple copies of DNA sequences. Four of the hybridization bands detected by a repetitive sequence probe, rTRS, were mapped to the ends of all the four chromosomes. Two or three of the bands detected by each of the other 12 probes were also mapped to different chromosomes. The bands detected by the same probe usually occurred in similar locations of different chromosomes. Loci detected by different DNA probes were often similarly arranged on different chromosomes. Chromosomes 8 and 9 showed colinearity of marker loci arrangement indicating a possible common origin. A segment on chromosome 9 was also very similar to the previously reported duplicated fragments on the ends of chromosomes 11 and 12 which were also detected in this study, indicating a likely common origin. Moreover, the various degrees of distributional similarity of the segments suggest a complex relationship among the chromosomes in the evolution of the rice genome. These results support the proposition that chromosome duplication and diversification may be a mechanism for the origin and evolution of the chromosomes in the rice genome.

Discriminant Classification Model of DNA Sequence

[Objective] The research aimed to construct the discriminant classification model of DNA sequence by combining with the biology knowledge and the mathematical method.[Method] According to the polarity nature of side chain radical in the amino acid,the classification information of amino acid which represented the sequence characteristic from the content and array situation of base was extracted from the different sequences that the amino acid content was different.The four-dimension vector was used to represent.Mahalanobis distance and Fisher discriminant methods were used to classify the given sequence.[Result] In the model,the back substitution rates of sample obtained by two kinds of classification methods were both 100%,and the consistent rate of classification was 90%.[Conclusion] In the model,the calculation method was simple,and the accuracy of classification result was higher.It was superior to the discriminant classification model which was only based on the base content.

Isolation and Characterization of Microsatellites in Snap Bean

The objectives of this study were to isolate and characterize microsatellites from a heat tolerant variety of snap bean (Phaseolus vulgaris L.) in order to generate polymorphic genetic markers linked to quantitative trait loci for heat tolerance. A genomic library contained 400-800 bp inserts was constructed and screened for the presence of (GA/CT) n and (CA/GT) n repeats. The proportion of positive clones yielded estimated of 3.72×10 4 such dinucleotide repeats per genome, roughly comparable to the abundance reported in other eukaryotic genomes. Twenty_six positive clones were sequenced. In contrast to mammalian genomes, the (GA/CT) n motif was much more abundant than the (CA/GT) n motif in these clones. The (GA/CT) n repeats also showed longer average repeat length (mean n =10.4 versus 6.5), suggesting that they are better candidates for yielding polymorphic genetic markers in the snap bean genome.

Isolation and Chromosomal Mapping of a Corn B Chromosome Specific RAPDs

B染色体存在于多种动植物中 ,具有很多独特的性状。B染色体与正常染色体在DNA组成方面十分相似 ,寻找B染色体特异序列一直是B染色体研究的难点和热点。通过对含有和不含有B染色体的两种玉米 (ZeamaysL .)基因组进行了RAPD分析 ,筛选到一个B染色体特异性分子标记B480。该标记与玉米的自主复制起始序列ARS1和ARS2同源 ,特别是该序列中的 2 5bp出现在多种模式生物基因组中。FISH的结果显示 。

DNA Molecular Identification of Botanical Origin in Chinese Herb Qingjiao

[Objective] The research aimed to distinguish Chinese herb Qingjiao from its botanical origin plants by comparing different DNA sequences,so as to provide a molecular basis for origin identification and quality evaluation.[Method] The cpDNA psbA-trnH and nrDNA ITS sequences of five Chinese herb Qingjiao plants,including Gentiana macrophylla pall.,Gentiana straminea Maxim.,Gentiana crassicaulis Duthie ex Burk.,Gentiana dahurica Fisch and Gentiana officinalis H.Smith,were amplified with PCR,and then sequenced by direct PCR sequencing method for homologous analysis.[Results] The length of cpDNA psbA-trnH of five plants was 316-318 bp;there were seven different haplotypes and seven variable sites;the GC content of the sequence was 21.2%;the phylogenetic clustering showed the same result as haplotype analysis.The length of nrDNA ITS sequence of five plants was 624-625 bp,there were five different haplotypes and 13 variable sites;the GC content of the sequence was 59.3%.The result of phylogenetic clustering suggested that G.dahurica and G.straminea,G.macrophylla and G.officinalis clustered together as sister clades,respectively.[Conclusion] The nucleotide differences of nrDNA ITS regions could be used for distinguishing botanical origin in Chinese herb Qingjiao.

Molecular Identification of Gallibacterium from Hen and Analysis of Its 16S rRNA Gene

Two strains of Gallibacterium, isolated from one laying hen flock in Zhengzhou City of Henan Province, were identified by the morphological observation, genus -specific PCR, and analysis of 16S rRNA gene, which was used to generate the phylogenetic tree, with the 21 members of the 12 genera belonging to Pasteurellaceae to analyze the homology. Two strains were named Yu-ZZ-HL-I-SLG and Yu-ZZ-HL-II-GZ. The comparative result of the 16S rDNA sequence shows that the 2 isolated strains are identical in sequence; the highest identity （99.9%） was observed between the isolated strain and one of the strains of Gallibacterium anatis （AF22.8002）, the homologies between the isolated strain and 3 strains of gallibacterium accessed in NCBI （AF22.8016, Gallibacterium genomosp. 1, AF228017, Gallibacterium genomosp. 2, AF22.8018, Gal- libacterium genomosp. 1 ） were above 97.1%, higher than that of the isolated strain and the other strains of the other 11 genera which were between 90.7% -93.2%. It can be seen from the phylogenetic tree that the 2 isolated strains and the other 4 strain of gallibacterium fell into the same branch, furthermore the 2 isolated strains and the strain of Gallibacterium anatis locate in an internal branch, indicating that the 2 isolated strains belong to Gallibacterium anatis.

Characteristic atom sequences of Nb-Mo alloys system in BCC structure and properties of disordered alloys

Comprehensively considering energy, volume and electronic structure of alloys, the ninth equation was determined as the interaction function of Nb-Mo alloys system in BCC structure on the basis of idea of systematic science of alloys, experimental lattice constants and heats of formation of disordered Nb（1-x）Mox alloys. The structural parameters and properties of Nb and Mo characteristic atoms sequences and corresponding characteristic crystals sequences were determined in Nb-Mo alloys system. The electronic structure and physical properties of disordered Nb（1-x）Mox alloys system were calculated according to concentration of characteristic atoms of disordered alloys. The change trend of physical properties is the same as that of electronic structure.

Sequence Analysis of Transcription Factor AtWRKY35 and Construction of Prokaryotic Expression Vector

As members of a super gene family, WRKY transcription factors are widely distributed in higher plants. ln this study, bioinformatic analysis of WRKY35, a member of the WRKY gene family, was carried out. Results indicated that tran-scription factor WRKY35 harbors a WRKYGQK core domain and a Cys2His2 or Cys2His/Cys zinc finger in the 5’ end without transmembrane domain. After PCR amplification and restriction digestion, WRKY35 gene fragment was ligated to prokaryotic expression vector PET28. This study provided basis for expression anal-ysis of WRKY35 protein and subsequent functional identification of WRKY35 gene.

Sequence and Molecular Evolution Analysis of Ubiquitin Proteins Encoded by Baculoviruses

[Objective] The aim of this study was to analyze the sequence characteristics and molecular evolution of ubiquitins encoded by baculoviruses.[Methods]Clustal W software was used for multiple sequence alignment analysis,and neighbor-joining method（NJ）and maximum parsimony method（MP）were used for the construction of phylogenetic tree.[Results]The baculoviral ubiquitins showed 73%-86% sequence identity to eukaryotic ubiquitin.Two heterogeneous regions of baculoviral ubiquitins were observed：one was the residues from 15-32,the other was located from residues 53 to 60.The else parts were conserved,where many functional amino acids were also observed.Phylogenetic analysis indicated that baculoviral ubiquitins could be divided into three sub-families,including sub-family GV,sub-family I and sub-family II.The molecular evolution of baculoviral ubiquitins might be under negative selection to maintain their functional and structural stability.[Conclusion]The analysis had provided reference for the researches on functional characterization of baculoviral ubiquitins.

Detection of Exogenous Gene Copies in Transgenic Soybean by Taqman Quantitative PCR Technique

[Objective] Taqman Quantitative PCR technique was adopted to detect the copies of exogenous nos terminator in transgenic hybrid soybean.[Method] With soybean Lectin as the endogenous reference gene,and gene complex DNA in non-GMO soybeans as the endogenous reference standard,the method of gradient dilution was used for separately calculate Ct value of endogenous reference gene and plasmid DNA and correlation standard curve equation of logarithm of copies,and then to calculate the copies of samples through substituting thus-obtained Ct into the standard curve equation.[Result] The standard curve equation of endogenous reference gene is y=-3.422x＋35.201,R2=0.998;and the standard curve equation of exogenous gene is y=-3.348x＋34.890,R2=0.999.Nos terminator and its lower boundary sequences in transgenic soybean is of single copy.[Conclusion] The study has provided a theoretical basis for determining exogenous gene copies in transgenic soybean.

Cloning and Sequence Analysis of ras Promoter from Auricuralia auricular Strain YBS-3

[Objective] The ras promoters were cloned from genomic DNA of Auricuralia auricular so as to provide promoters for breeding better species by genetic technology. [Method] PCR was used to clone promoters with the genomic DNA of A. auricular strain YBS-3 as template,and then the sequences of four ras promoters were obtained. The analysis of promoter sequences was made by three promoter analysis software：Promoter prediction,Place and TFSEARCH ver.1.3. [Result] Four fragments contained core elements of promoter including TATA-box and CAAT-box,and many other important cis-elements such as GATA BOX,GCGC BOX,CCAAT BOX1,etc. At the same time,there was at least one transcriptional start site in these sequences. And several transcription factor binding sites were detected in these sequences. [Conclusion] Four promoter fragments all had promoter function theoretically.

Correlation-based subblock partition design for PTS approach

In order to exploit the capability of the peak-to-average power ratio（PAPR）reduction afforded by the partial transmit sequences （PTS）approach in orthogonal frequency division multiplexing（OFDM）systems, subblock partition schemes for the PTS approach are studied. The motivation is to establish the relationship between the subblock partition and the capability of PAPR reduction through the periodic autocorrelation functions （ACFs）of partial transmit sequences and the periodic cross-correlation functions（CCFs）of signal candidates.Let Q represent the variation of the square magnitudes of ACFs.It is found that the lower the Q-value is, the better PAPR performance can be achieved, which is introduced as a design criterion for subblock partition.Based on this criterion, four common partition methods are compared and an efficient partition strategy is proposed. It is shown that structured partition schemes with low computational complexity have a large Q-value, leading to a poor PAPR performance.The new strategy can be regarded as a trade-off between PAPR performance and computational complexity.The simulation results show that the strategy can achieve an optimal performance with a relatively low complexity and, moreover,does not increase the amount of side information.

Cloning and Expression Patterns of a Metallothionein-like GenehtMT2 of Helianthus tuberosus

A novel cDNA sequencehtMT2, which encodes a type 2 metallothionein_like protein, was isolated from Helianthus tuberosus L. tuber cDNA library. The whole sequence is 509 bp, including an open reading frame (ORF) of 240 bp, a 5′ UTR of 62 bp and a 3′ UTR of 207 bp. Two genomic sequences covering the coding region ofhtMT2were cloned by PCR reaction. Sequence analysis revealed that the genomic sequences htMTG_1 of 986 bp and htMTG_2 of 982 bp were both composed of three exons and two introns. The deduced protein consisted of 79 amino acid residues with a predicted molecular weight of 7.8 ku (kD). Amino_terminal and carboxy_terminal domains contained 8 and 7 cysteine residues respectively, separated by a central cysteine free spacer. Sequence alignment revealed that the predicted protein ofhtMT2 was homologous to type 2 metallothioneins (MTs) of plants. Southern blotting analysis indicated that htMT2was encoded by a small multi_gene family in H. tuberosus genome. Northern blotting analysis showed that htMT2 transcripts were detected in stems, leaves and leafstalks, but no transcripts were detected in roots. The expression level in stems was the highest among the above tissues. Transcripts in stems were significantly reduced by Cu 2+ treatment. Judging from the homologies between the deduced HtMT2 and other type 2 plant metallothioneins as well as responses to metal ions, we believe thatwere cloned by PCR reaction. Sequence analysis revealed that the genomic sequences htMTG_1 of 986 bp and htMTG_2 of 982 bp were both composed of three exons and two introns. The deduced protein consisted of 79 amino acid residues with a predicted molecular weight of 7.8 ku (kD). Amino_terminal and carboxy_terminal domains contained 8 and 7 cysteine residues respectively, separated by a central cysteine free spacer. Sequence alignment revealed that the predicted protein ofhtMT2 was homologous to type 2 metallothioneins (MTs) of plants. Southern blotting analysis indicated that htMT2was encoded by a small multi_gene family in H. tuberosus genome. Northern blotting analysis showed that htMT2 transcripts were detected in stems, leaves and leafstalks, but no transcripts were detected in roots. The expression level in stems was the highest among the above tissues. Transcripts in stems were significantly reduced by Cu 2+ treatment. Judging from the homologies between the deduced HtMT2 and other type 2 plant metallothioneins as well as responses to metal ions, we believe that[ShtMT2 encodes a new type 2 metallothionein.

Molecular Characterization of Four ADF Genes Differentially Expressed in Cotton

Actin depolymerizing factor （ADF）, highly conserved in all eukaryotic cells, is a low molecular mass of actin-binding protein, which plays a key role in modulating the polymerizing and depolymerizing of the actin filaments. Four cDNAs （designated GhADF2, GhADF3, GhADF4, and GhADF5, respectively） encoding ADF proteins were isolated from cotton （Gossypium hirsutum） fiber cDNA library. GhADF2 cDNA is 705 bp in length and deduces a protein with 139 amino acids. GhADF3 cDNA is 819 bp in length and encodes a protein of 139 amino acids. GhADF4 cDNA is 804 bp in length and deduces a protein with 143 amino acids. GhADF5 cDNA is 644 bp in length and encodes a protein of 141 amino acids. The molecular evolutionary relationship of these genes was analyzed by means of bioinformatics. GhADF2 is closely related to GhADF3 （99% identity） and PetADF2 （89% identity）. GhADF4 is closely related to AtADF6 （78% identity）, and GhADF5 is closely related to AtADF5 （83% identity）. These results demonstrated that the plant ADF genes are highly conserved in structure. RT-PCR analysis showed that GhADF2 is predominantly expressed in fiber, whereas, GhADF5 is mainly expressed in cotyledons. On the other hand, it seems that GhADF3 and GhADF4 have no tissue specificity. Expression levels of different ADF genes may vary considerably in the same cell type, suggesting that they might be involved in regulating tissue development of cotton and the each ADF isoform may diverge to form the functional difference from the other ADFs during evolution.

The σ-duals of Sequence Spaces

In this paper, the σ_duals of two classes important sequence spaces l 1(X) and l ∞(X) are investigated, and shows that some topology properties of locally convex space (X,τ) can be characterized by the σ _duals. The criterions of bounded sets in l 1(X) and l ∞(X ) with respect to the weak topologies generated by the σ _duals are obtained. Furthermore, a Schur type result and an automatic continuity theorem of matrix transformation are established.

Molecular Cloning and Sequence Analysis ofBoPGIP2 Gene from Brassica oleracea L. var. alboglabra

PGIP gene was obtained from Brassica oleracea L. var. alboglabra, named BoPGIP2. The full length of BoPGIP2 gene is 1 102 bp and the exon is 993 bp which encodes a protein of 330 amino acids with a predicted molecular mass of 37.1 kDa, interrupted by one intron of 95 bp in, length. Sequence analysis revealed that it has five potential N-giycosylation sites, two protein kinase C phosphrylation sites, five casin kinase Ⅱ phosphrylation sites and four N-myristoylation sites. The amino acids sequences alignment confirmed that ^145 LRR stucture was highly conserved in all aligned PGIP sequences.