昆虫学报2012，Vol.55Issue(3)：259-266,8.
黑腹果蝇GABA受体GRD亚基和LCCH3亚基的克隆、表达与鉴定
Cloning, expression and identification of GRD and LCCH3 subunits of GABA receptor in Drosophila melanogaster
摘要
Abstract
γ-Aminobutyric acid receptor (GABAR) is one of the most important insecticide targets. In this study, the fragments of genes of GRD and LCCH3 subunits in Drosophila melanogaster were amplified by RT-PCR and cloned into pET-32a expression vector. Homologous analysis showed that they have more than 99% identity with the published amino acid sequences of the homologous genes in GenBank, without frameshift mutation. The recombinant plasmids were transformed into Escherichia coli and expressed under IPTG induction. LCCH3 gene was expressed successfully, but GRD gene failed. The expressed LCCH3 subunit was purified by the processes of inclusion body washing, denaturation, Ni2+ affinity chromatography and renaturation. The secondary structure of LCCH3 subunit was determined by circular dichroism (CD) spectroscopy, the result showed that this subunit was rich in β-strand. The results provide important information for the study of relationship between the structure and function of GABA receptor.关键词
黑腹果蝇/GABA受体/GRD亚基/LCCH3亚基/基因克隆/表达/鉴定Key words
Drosophila melanogaster/ GABA receptor/ GRD subunit/ LCCH3 subunit/ gene cloning/ expression/ identification分类
生物科学引用本文复制引用
张晓敏,马彩霞,任天瑞,王丰..黑腹果蝇GABA受体GRD亚基和LCCH3亚基的克隆、表达与鉴定[J].昆虫学报,2012,55(3):259-266,8.基金项目
国家自然科学基金项目(20872093, 21172147) （20872093, 21172147）
上海市教委基金项目(11ZZ122) （11ZZ122）
