旋毛虫新生幼虫cDNA文库的构建与鉴定

利用λZAPExpress载体成功地构建了中国猪株旋毛虫分离株Trichinellaspiralis新生幼虫的cDNA文库 ,并对其重组噬菌体质粒pBK—CMV进行酶切鉴定 .结果表明 :所构建的cDNA文库容量为 1 92× 10 6,重组效率为 98 6 % ,所有的克隆片段都在 0 5— 2 0Kb之间 ,说明此cDNA文库几乎覆盖了全部mRNA .     

青岛文昌鱼profilin样蛋白基因的克隆与同源性分析

对青岛文昌鱼神经胚cDNA进行序测，获得了6个profilin样蛋白基因的EST，经拼接得到编码文昌鱼profilin样蛋白基因的cDNA序列并据此演绎了文昌鱼profilin样蛋白的氨基酸序列。对演绎的文昌鱼profilin样蛋白的一级结构进行了分析，对其二级结构进行了预测，并将其与多种真核生物中的profilin进行同源性比较，为profilin分子进化的研究提供资料。     

炭疽毒素及其克隆受体的研究进展

综述炭疽毒素研究的最新进展，炭疽毒素由3种蛋白组成：致死因子（L ethal factor,LF)，水肿因子(edema factor,EF)和保护性抗原（protective antigen,PA)，本文主要讲述了炭疽毒素的关键致病因子－致死因子（LF）的结构和功能，炭疽素受体（anthrax toxin receptor,ATR）的结构及其特征性功能基团，介绍了通过基因互补对ATR进行克隆的方法，并讨论了ATR和ATR的cDNA克隆在炭疽治疗的可能应用。     

单细胞克隆法快速筛选重组昆虫杆状病毒

首先采用显微操作系统挑取包含有重组病毒的单细胞，成功地获得了重组杆状病毒的单细胞克隆，并得到了重组病毒的纯培养，对有多角体的和无多角体的重组病毒细胞的挑取表明，这一方法对重组杆状病毒的筛选具有普遍适用性，我们发展的这一方法比常规的空斑技术简便、便速，只需两周左右即完成重组病毒的筛选。     

2.15 kDa人脑髓鞘硷性蛋白cDNA的体外扩增

髓鞘硷性蛋白(MBP)是脊椎动物神经系统髓鞘膜的外周蛋白,它对有鞘神经的绝缘和块速传导具有重要作用.人和鼠MBP基因均含7个(Ⅰ—Ⅶ)外显子,但其原始转录体剪接方式不同.人MBP至少有21.5,20,18.5和17kDa等4种,其中唯21.5kDa是     

从微量全血直接进行反转录-聚合酶链反应

在转录水平上研究基因表达及调控,通常要求制备纯度高、未降解的总RNA。提取总RNA的方法一般较复杂。Kawasaki描述过一个从1—2ml全血制备RNA的方法。本文报道了一个更为简便的从微量全血不经抽提RNA直接进行RT-PCR的方法。即将2—20μ1全血低渗破红细胞,离心得到白细胞沉淀,在有RNA酶抑制剂存在的条件下再低渗使白细胞破坏,释放出RNA。此液直接用于RT—PCR,并与用AGPC法抽提的总RNA为     

Cloning and roles of goldfish maternal factor β-Catenin cDNA in embryonic development

Interaction between nucleus and cytoplasm has been focused in the field of animal embryonic development, in which study of maternal factors is required positively. β-Catenin, an important maternal factor in early embryogenesis, has been analyzed in its expression pattern and functions in this paper. We have cloned goldfish β-Catenin cDNA gene and compared it with zebrafish β-Catenin cDNA. High homology was found in cDNA and in amino acid sequences between them, 93% (2227/2384 bp) and 98.5% (768/780 aa) respectively. The expression pattern of β-Catenin by in situ hybridization and the roles of β-Catenin on embryonic development by co-injection of anti-sense RNA and reporter gene, EGFP have been investigated in the whole process of goldfish embryonic development. The results suggest that β-Catenin presents dynamic distribution, mainly locates at body axis, dorsal tissues, head and tail structures after being fertilized. The loss of β-Catenin activity would cause serious destruction of embryo in dorsal tissues and in anteroposterior axes, and leads embryos to die before larva get hatched.     

Infectious foot-and-mouth disease virus derived from a cloned full-length cDNA of OH／CHA／99

The China foot-and-mouth virus (FMDV) isolate OH/CHA/99 was isolated from swine, which was unable to infect bovine thyroid cells in vitro or to cause typical disease in bovines following intradermal inoculation in the tongue. To enhance antigenicity, replication, maturation and pathogenicity studies of OH/CHA/99, an infectious fulllength cDNA clone, designated pBIFMDV, was prepared. The in vitro and in vivo biological properties of the virus derived from pBIFMDV were studied by analyzing antigenicity, plaque morphology and virulence in pigs. The results showed that the virus derived from pBIFMDV had the same biological properties as the parent strain OH/CHA/99; the fulllength infections cDNA clone, pBIFMDV, will be very useful in studies of the antigenicity, virulence, pathogenesis, maturation and replication of FMDV.