家蚕内网虫属微孢虫核糖体小亚单位RNA(SSUrRNA)核心序列的克隆和序列分析

Endoreticulatus bombycis is a new pathogenic microspordia isolated from the silkworm larvae.With polymerase chain reaction(PCR) method,we amplified a fragment of the core sequence of the small subunit ribosomal RNA( SSUr-RNA) of Endoreticulatus bombycis.The SSUrRNA fragment was inserted into pMD18-T Vector and then cloned.It had a length of 1230 nucleotides and a percentage GC content of 51.3%.The accession number in Genbank is gill 181769|AY009115.The secondary structure model of the SSUrRNA of Endoreticulatus bombycis was constructed both on the bases of the sequence alignment and RNAFOLD program of the PC-GENE package.The secondary structure model revealed that Endoreticulatus bombycis lacked many cukaryotic hclices.such as heli10,helix 11.helix 18,helix 43 and helix 46,but it has V4 region and has more of prokaryotic character.Analyzed by Blast,Endoreticulatus bombycis.Endoreticulatus schubergi and pleistophora sp.(Sd-Nu-IW8201) have a high similarity,over 98%,Phylogeny tree of Endoreticulatus and Pleistophora species showed that Endoreticulatus and Pleistophora sp.(Sd0Nu-IW8201) was in a group and the other Pleistophora species were in another groupActa Zoologica Sinica 49(3):414-420,2001].

Cloning and analysis of the core sequence of small subunit ribosomal RNA of Endoreticulatus bombycis

Endoreticulatus bombycis is a new pathogenic microspordia isolated from the silkworm larvae. With polymerase chain reaction (PCR) method,we amplified a fragment of the core sequence of the small subunit ribosomal RNA (SSUrRNA) of Endoreticulatus bombycis. The SSUrRNA fragment was inserted into pMD18-T Vector and then cloned. It had a length of 1 230 nucleotides and a percentage GC content of 51.3%. The accession number in GenBank is gi11181769|AY009115. The secondary structure model of the SSUrRNA of Endoreticulatus bombycis was constructed both on the bases of the sequence alignment and RNAFOLD program of the PC-GENE package. The secondary structure model revealed that Endoreticulatus bombycis lacked many eukaryotic helices, such as helix10, helix 11, helix 18, helix 43 and helix 46,but it has V4 region and has more of prokaryotic character. Analyzed by Blast, Endoreticulatus bombycis, Endoreticulatus schubergi and Pleistophora sp.(Sd-Nu-IW8201) have a high similarity, over 98%. Phylogeny tree of Endoreticulatus and Pleistophora species showed that Endoreticulatus and Pleistophora sp.(Sd-Nu-IW8201) was in a group and the other Pleistophora species were in another group .