日本沼虾感光器Gqα基因的扩增及分析

Molecular cloning and analysis of Gq proteinαsubunit gene in Maacrobroachium nipponense Photoreceptor

采用分子生物学方法,提取日本沼虾感光器的总RNA,反转录成c DNA．根据Gq蛋白α亚基的保守序列设计简并引物,通过巢式PCR扩增出日本沼虾感光器Gq仪基因的片段,并将该片段克隆到pMD 18-T载体上进行序列测定,结果显示此基因片段由369bp组成．翻译成氨基酸序列(123aa),Blast搜索结果比较该片段氨基酸序列与美洲鲎、美洲龙虾、冈比亚按蚊等6种动物的Gqot氨基酸保守序列具有高度同源性(83．74%～95%),其中与冈比亚按蚊的Gqa氨基酸序列同源性最高(95%)．

The total RNA in the photoreceptor of Maacrobroachium nipponense was separated and reversed to cDNA by RT-PCR. The specific primers had been designed according to the conservative amino acids sequence of Gqα-encoding genes for the purpose of amplification of similar Gqα - encoding genes from the genome ofMaacrobroachium nipponense photoreceptor. One DNA fragment had been obtained by nest PCR , cloned into pMD18 -T vector and sequenced. Sequence analysis showed the amplifying fragment was composed of 369bp which can be translated to 123 amino residues. The deduced amino acid sequence of the amplifing fragment showed high homology compared with the amino acids sequence of Gq protein α subunit in other six species from Genbank（ 83.74% -95 % ）by BLAST revealed;the most high homology is 95 % between Maacrobroachium nipponense and Anopheles gambiae str. Based on PCR properties and molecular weights, it was deduced that the amplifing fragment is Gqα-encoding genes in the photoreceptor of Maacrobroachium nipponense.