菜粉蝶微孢子虫核糖体RNA(rRNA)编码基因的研究

用PCR方法扩增、克隆了菜粉蝶微孢子虫核糖体小亚单位RNA(SSUrRNA)编码基因的核心序列 1 2 0 5bp后 ,进一步克隆到菜粉蝶微孢子虫SSUrRNA基因 3′端至LSUrRNA基因 5′端 (580R区 ) 657bp长的序列。与GenBank中对应序列比较后 ,在 657bp这段序列鉴定出菜粉蝶微孢子虫SSUrRNA基因 3′末端、rRNA基因内转录间隔区 (ITS)及LSUrRNA基因 5′端 (580R区 ) ,它们分别位于该序列中 1 45位、1 46 1 86位及 1 87位。与SSUrRNA基因核心序列拼接后SSUrRNA全基因长为 1 2 4 5bp ,rRNA基因内转录间隔区为 41bp及核糖体大亚单位RNA(LSUr RNA)编码基因 580R区为 470bp。同时还构建了菜粉蝶微孢子虫SSUrRNA的完整二级结构。关于微孢子虫rRNA基因的克隆及SSUrRNA的二级结构在国内尚属首次报道 ,它为进一步利用核糖体RNA编码基因及SSUrRNA的二级结构对不同微孢子虫的分类及亲缘关系的确定奠定了基础

STUDIES ON THE RIBOSOMAL RNA GENE(rDNA)OF A MICROSPORIDIUM ISOLATED FROM PIERIS RAPAE L

Nuclotide sequence (1205 bp) of small subunit ribosomal RNA (SSUrDNA) of a microsporidium isolated from Pieris rapae L. (abbr:MPr) was specifically amplified by polymerase chain reaction (PCR). Another fragment of 657 bp downstream of MPr SSUrDNA 3'end was amplified with two other primers. Within this 657 bp fragment, the putative 3'terminus of MPr SSUrDNA and the extreme 5' of large subunit ribosomal RNA gene. (LSUrDNA) were identified, which situated at base 145, 146-186 and 187, respectively. Then the full sequence of MPr ssurDNA is 1245 bp. Its GC content was also nearly 34%. The ITS region (internal transcribed spacer), positioned between the ssu and LSUrRNA genes, was found to be 41 bp in length. The LSUrDNA 580r region of MPr is 470 bp, longer than 437 bp of Nosema apis, 447 bp of Nosema algerae. The secondary structure of MPr SSUrRNA was constructed. These analyses of MPr rRNA gene contributed to the somewhat limited microsporidian taxonomic classification based on morphology.