拟南芥雄性不育突变体ms1521的基因定位

ms1521是经EMS诱变筛选得到的一拟南芥雄性不育突变体,通过背景纯化与遗传分析,发现ms1521突变体是受隐性单基因控制,形态学观察表明：突变体的花缺失部分花瓣,雄蕊比较短,花药肥大,部分雄蕊的花药成丝状,利用图位克隆的方法对不育基因MS1521进行了定位,结果表明：MS1521位于第一条染色体分子标记F17F8Alu1和T19E23之间161kb的区间内,数据库预测,其中有11个与花发育有关的基因,试验将有助于对目的基因的克隆,控制花器官研究和雄性不育分子机制的研究。     

粗糙脉孢菌BenR基因在哈茨木霉中的定点转化

以β2-微管蛋白基因为定点整合位点,通过原生质体法将多菌灵抗性基因转化到哈茨木霉中,获得了具有多菌灵抗性的生物防治工程菌株．多菌灵抑制抗性转化子菌丝生长的ECso值达471．26μg／mL,比哈茨木霉原菌株提高1200倍以上;转化子对多菌灵的抗性具有遗传稳定性,且在无选择压力下菌丝生长速度及菌落形态与原菌株无显著差别;抑菌活性检测结果表明,3个转化子对立枯丝核菌均具有较强的抑菌活性,对菌丝生长的抑制率分别为87．5％、86．3％和85％．     

Efficient transformation and expression of gfp gene in the edible mushroom Pleurotus nebrodensis

An efficient transformation method mediated by PEG-protoplasts was developed for the newly commercial edible mushroom Pleurotus nebrodensis. Two plasmids were used to co-transform protoplasts of P. nebrodensis. One plasmid is pAN7-1 containing a positive selectable marker gene hph conferring hygromycin B resistance. Another plasmid is pBlue-GFP containing a reporter gene gfp conferring green fluorescent protein. PCR and Southern blot analysis showed that hph gene or/and gfp gene were integrated into the genome of P. nebrodensis transformants. The transformation efficiency of the positive selectable marker gene hph was 3 transformants per microgram of plasmid pAN7-1 DNA, which was about 30 times higher than that previously reported in thoroughly studied Pleurotus species such as Pleurotus ostreatus. The transformation efficiency of the reporter gene gfp was 9 transformants per microgram of plasmid pBlu-GFP DNA. The co-transformation efficiency was 23.68%. This is the first report that a ＂reporter＂ gene, green fluorescent protein gene can be successfully stably exoressed in this Pleurotus species.     

Characterization of codon usage bias in the dUTPase gene of duck enteritis virus

A comparative analysis of the codon usage bias in the newly discovered dUTPase gene （Assigned Accession No.： DQ486149） of the duck enteritis virus （DEV） and the dUTPase gene of 32 reference herpesviruses was performed. The results indicated that the DEV dUTPase gene encodes a protein of 477 amino acids, which includes five conserved motifs with a 3-1-2-4-5 arrangement. The codon adaptation index （CAI）, effective number of codons （ENC）, and GC3s values indicated synonymous codon usage bias in the dUTPase gene of herpesviruses, and this synonymous bias was correlated with host evolution. The codon usage patterns of the DEV dUTPase gene were phylogenetically conserved and similar to that of the dUTPase genes of the avian alphaherpesvirus. Although codon usage in each microorganism was different, there were no strain-specific differences among them. Sixty-one codons in the predicted polypeptide, with a strong bias towards A and T at the third codon position, were used. Comparison of the codon usage in the dUTPase gene of different organisms revealed that there were 19 codons showing distinct codon usage differences between the DEV and Escherichia coli dUTPase genes; 16 between the DEV and yeast dUTPase genes; and 15 between the DEV and human dUTPase genes. Analysis of variance （ANOVA） showed significant differences between the DEV and yeast dUTPase genes （r = 0.536, P 〈 0.01）. The extent of codon usage bias in the DEV dUTPase gene was highly correlated with the gene expression level, therefore the results may provide useful information for gene classification and functional studies.     

矮秆种质的赤霉酸反应和株高遗传模型的分析

通过赤霉酸处理及遗传模型分析小麦／冰草衍生系的矮秆种质得知：8个矮秆种质均为赤霉酸不敏感型。其中的3648—3652的株高是多基因控制的数量性状,其遗传模型是两对主基因加多对微效基因控制的混合遗传模型。     

Cloning, Characterization, and Expression Analysis of Calreticulin from Pearl Oyster Pinctada fucata

Calreticulin is a unique calcium-binding protein with multiple functions mostly located in the sarcoplasmic/endoplasmic reticulum. A large amount of calcium is absorbed from the medium and transported to mineralization sites during biomineralization in pearl oyster. This paper describes the cloning of the full-length cDNA of calreticulin from Pinctada fucata, namely PCRT. PCRT encodes a deduced 414-amino acid protein, which includes a predicted 17- amino acid signal peptide and an endoplasmic reticulum retrieval sequence HDEL. The protein shows 63%-76% sequence identity and shares some common characteristics with calreticulins from other species. Semi-quantitative RT-PCR indicates that PCRT is ubiquitously expressed in all tissues tested with the highest expression in the hemolymph and the mantle. In situ hybridization analysis of PCRT in the mantle showed strong signals in the inner fold, the inner side of middle fold, and the inner side of outer fold of the mantle epithelium, All these results suggest PCRT might be involved in Ca^2＋ transport and storage during oyster biomineralization.     

犬细小病毒CPV-SHZ分离株VP2基因的克隆与序列分析

以本实验室分离鉴定犬细小病毒新疆石河子株（CPV-SHZ）的DNA为模板,根据基因库已发表CPV序列设计合成了VP2基因的1对特异引物,进行聚合酶链式反应,扩增出约1.7kb的片段,按常规方法克隆进pMD18-T载体,经EcoR I和Sal I双酶切筛选到阳性质粒。测序得到VP2全基因组序列,并登陆Genbank（EU170352）。进一步对该片段进行序列分析,结果表明：所扩增基因片段长度为1755bp,与CPV参考株毒株V154（Type2a）、LCPV-V204（Type2b）、LCPV-V139（Type 2c（a））、LCPV-V203（Type 2c（a））,其核苷酸的同源性分别为99.32%、98.75%、98.97%、98.69%,确定CPV-SHZ株基因型为2a型,将其同我国和世界其他国家主要分离株进行基因系统发生进化关系分析,结果表明,其与我国北京分离株BJ018/07亲缘关系较近。     

植物无标记转化的诱导表达Cre/loxP重组系统的构建

应用Cre/loxP系统位点专一性重组的特点构建诱导表达的定位重组系统,用以特异性的敲除转基因植物的标记基因.为了获得诱导表达启动子,从大豆基因组DNA中用pfu酶克隆热激蛋白启动子gmhsp17.5c,将其克隆到pUC118-HincⅡ载体并测序.结果表明,508nt的gmhsp17.5c与已报道序列(GenBank,AF544399)比较,核苷酸的同源性为99.8%.利用该诱导启动子分别构建了含gmhs p17.5c-cre基因组件和gmhsp17.5c-gus基因组的诱导型植物表达载体pC23HC和pC23HG.此外1个含有loxP-gus-loxP组件的组成型植物表达载体pC23LG被构建.通过对3个植物表达载体做多重酶切及亚克隆后测序分析表明载体pC23HC全长10947bp,载体pC23HG全长11396bp,载体pC23LG全长11900bp,符合预期设计.一套由pC23HC和pC23LG组成的植物无标记转化的诱导表达Cre/loxP重组系统被构建,为进一步将诱导表达的标记基因删除系统用于植物的无标记转化奠定基础.     

鸭瘟病毒强、弱毒株TK基因的克隆与序列测定

根据GenBank上已发表的鸭瘟病毒TK基因核苷酸序列,设计并合成了1对引物,分别扩增鸭瘟病毒标准强毒株(DPV-F34)和鸭瘟鸡胚化弱毒疫苗株(C-KCE)的TK基因,将它们分别克隆入pBS-T载体,转化TOP10大肠杆菌,对阳性的重组质粒进行序列测定.结果表明:DPV-F34与DPVC-KCE的TK基因长度均为1077bp,编码358个氨基酸,二者的核苷酸与氨基酸组成具有很高的同源性,分别为99.4%和98.9%.