一个RNAi载体上反向重复片段的测序策略

相邻的反向重复DNA片段有形成单链内二级结构的倾向,属于一种测序困难的DNA模板。解决RNAi载体插入的反向重复片段的测序问题,为该类载体正确性的测序验证奠定基础。采用常规分子克隆方法构建表达小麦TaATG2串联反向重复片段的RNAi载体,设计2种策略对经菌落PCR初步鉴定的载体进行测序验证:一种是以完整的载体质粒为模板进行测序;另一种是先对载体进行酶切处理,切除反向重复片段中的一个后对保留另一个片段的线性载体进行测序。结果表明,第一种测序策略受到串联反向重复片段形成的单链内部二级结构的影响,测序信号在反向重复片段处出现衰减或乱峰,无法读取序列。第二种测序策略排除了2个反向重复片段之间的干扰,保留在载体上的片段测序信号清晰,序列准确。采用酶切切除一个片段后进行测序的方法,经过2次酶切和2次测序可以有效地对载体上的2个反向重复片段分别进行序列测定,进而确认构建载体的正确性。     

建立以TK基因为标记将外源基因导入臭曲霉（Asper—gillus...

A new system for selection of transformed Aspergillus foetidus was reported. In this system, TK- A. foetidus which were constructed by homologous recombination of mutated TK gene of vaccinia virus with TK gene of A. foetidus were screened by adding BUdR in agar plates. Conditions for screen of TK+ A. foetidus strain, transformation of A. foetidus and selection of transformed TK- A. foetidus have been studied. By using this system, several transformed A. foetidus which contained HBsAg gene derived bf a promoter H8 cloned from genomic DNA of A. foetidus were isolated. It was demonstrated that HBsAg gene was integrated into the chromosome DNA of A. foetidus by Southern blot after many passages of spores. ELISA showed that HBsAg was positive in the growth medium (p/n = 20). The 22 nm particles which were very similar to the HBsAg particles in human serum were found in the growth medium by immunoelectromicroscope. Western blot also gave the specific bands. All these data showed that HBsAg gene was expressed in A. foetidus and the products were secreted into the growth medium. The selection system using TK gene as marker could generally be used to study the expression of foreign gene in A. foetidus.     

蒸腾抑制剂对组培苗出瓶移栽后成活率的影响

蒸腾抑制剂０．１％ＣａＣｌ２和０．１％ＦＡ旱地龙可提高培苗出瓶移栽的成活率，对成活率低的Ⅱ、Ⅲ级组培苗效果相对较显著。前者宜在移栽后即刻喷施，后者宜在移栽前４ｄ于瓶中喷施。     

异域作用因子

基因表达时基因携带的遗传信息从世代相传的遗传物质DNA上通过转录传递到RNA，然后由RNA指导合成蛋白质，从而表现出基因的生物活性。基因的表达受环境因素的影响，并受生物体的控制。真核生物的基因表达在不同水平上受到调控。在细胞核中有转录时的调控，它决定基因是否能被转     

略谈真菌和植物之间的相互关系

真菌曾被列入植物的范畴，被作为植物学的一个分支学科的研究对象，它们被看作一类不含叶绿体的植物。目前已经了解，在地球上真菌的种类在１００万种以上，而维管植物的种类在３０－３５万种之间。因此，研究和了解地球上这两组庞大的生物群体之间的相互关系是十分有意义的，也是目前非常活跃的科学领域。真菌和植物之间的相互作用，有相互依赖，也即相生的一面。现代化石和分子方面的证据证实，真菌和植物的这种相互作用是从远古开始的，是从这二组生物群体出现在陆地上就开始的。事实上，学者认为，最远古、最原始的植物可能是借助于同真…