九种微孢子虫核糖体小亚单位RNA(SSUrRNA)基因拷贝数的研究  

Analyses of the Copies of SSUrRNA Genes of Nine Microsporidia Pathogenic to the Silkworm,Bombyx mori

作　　者：王见杨[1] 黄可威[1] 赵昀[2] 陆长德[2] 

机构地区：[1]中国农业科学院蚕业研究所,镇江212018 [2]中国科学院上海生物化学研究所

出　　处：《蚕业科学》2001年第3期200-205,共6页ACTA SERICOLOGICA SINICA

基　　金：国家"九五"重点科技攻关课题96 -6 16 -0 2 -0 3

摘　　要：以已克隆测序的家蚕微孢子虫 (镇江株 ,Nosemabombycis)的核糖体小亚单位RNA(SSUrRNA)编码基因(12 0 5bp)为模板 ,用随机引物合成法标记的探针 ,在与 9种微孢子虫及家蚕基因组DNA的 6种限制性内切酶的酶切产物的Southern杂交图谱中 ,9种微孢子虫基因组DNA酶切产物的杂交图谱极为相似 ,而与家蚕基因组DNA的任何一种酶的酶切产物均无杂交信号 ,表明微孢子虫和家蚕的SSUrRNA基因同源性很低或没有同源性。同时还证实了已克隆的SSUrRNA基因来源于微孢子虫基因组DNA ,不是从家蚕基因组DNA扩增而来。在酶解较为完全的酶切产物的杂交图谱中 ,微孢子虫基因组DNA中SSUrRNA基因的拷贝数至少在The α 32 P radiolabelled probe was synthesized by random oligonuclotide primers using Nosema bombycis (Zhenjiang strain)SSUrRNA gene(1 205 bp)cloned as template.It was used to probe Southern blot of Eco RⅠ,Hin dⅢ, KpnⅠ,PstⅠ,PvuⅡ,XhoⅠ ～digested genomic DNA from each of the nine microsporidia isolates and the host( Bombyx mori ).The hybridizing maps in nine microsporidia genomic DNA are very similar to each other,indicating the nine microsporidia are very closely related to each other in phylogenetic relationship.But there was no hybridizing band in any enzyme～digested host genomic DNA.It is suggested that there was low/no homology between SSUrRNA genes of microsporidia and host.At the same time,it also confirmed that the cloned SSUrRNA gene was of microsporidia origin.Southern blot analysis revealed that there were at least 15 hybridizing bands in nearly completely enzyme～digested microsporidia genomic DNA

关 键 词：微孢子虫 SSUrRNA基因 拷贝数 核糖体小亚单位RNA 基因克隆 家蚕 

分 类 号：S884[农业科学—特种经济动物饲养]