大眼长蝽卵黄原蛋白基因克隆、序列分析及表达研究

doi:10.13560/j.cnki.biotech.bull.1985.2015.10.024

摘要/Abstract

摘要： 旨在为研究大眼长蝽（Geocoris pallidipennis）卵黄原蛋白（Vitellogenin, Vg）分子特性及其基因生理功能。利用RT-PCR、RACE及ELISA方法对大眼长蝽Vg基因进行克隆、序列分析和表达研究。该基因cDNA全长5 667 bp（GenBank登录号：KP688587）, 编码1 848个氨基酸残基, N-末端的前19个氨基酸为信号肽。氨基酸序列中有两个保守的多聚丝氨酸区域和RXXR酶切位点, 接近C-末端有GLAG基序, 其后有5个保守的半胱氨酸残基, DGYR基序位于GLAG上游18个氨基酸残基处。该基因编码氨基酸序列与其它半翅目昆虫Vg氨基酸序列相似度较高, 氨基酸序列分析显示它有Vg的典型特征, 表明克隆的cDNA序列是大眼长蝽的Vg基因序列。ELISA检测发现随着发育时间的延长, 卵黄原蛋白表达量逐渐增加, 羽化后22 d达到高峰, 随后开始下降, 结果表明大眼长蝽雌虫卵黄原蛋白的表达量与大眼长蝽产卵量紧密相关。

关键词: 大眼长蝽, 卵黄原蛋白, Vg基因, 序列分析, Vg表达

Abstract: It was to study Vitellogenin（Vg）and the function of Vg gene in G. pallidipennis. In this study, the gene cloning, sequence analysis and expression studies of Vg gene of G. pallidipennis were carried out by RT-PCR, RACE and ELISA. The full-length cDNA of the Vg gene was 5 667 bp（GenBank accession number：KP688587）, encoding 1848 amino acids residues, and there was a signal peptide of 19 amino acids in N-terminal. There were 2 conserved polyserine regions and 1 RXXR cleavage site in amino acid sequences. Close to the C-terminus there was a GLAG motif followed by 5 conserved cysteine residues, and a DGYR motif was located in the 18th residue of the GLAG upstream. The deduced amino acid sequence of the Vg gene showed a high similarity with the Vg sequences from other hemipterainsects. Typical features of Vg were found through sequence analysis. The results indicated that the cDNA obtained was Vg gene from G. pallidipennis. The Vg detected by ELISA showed it increased gradually and reached the peak on the 22th day after adult emergence, then decreased. The results also indicated that the egg production was closely correlated to Vg expression.

Key words: Geocoris pallidipennis, vitellogenin, gene cloning of Vg, sequence analysis, expression of Vg

梁慧芳,曾凡荣,毛建军. 大眼长蝽卵黄原蛋白基因克隆、序列分析及表达研究[J]. 生物技术通报, 2015, 31(10): 149-156.

Liang Huifang, Zeng Fanrong, Mao Jianjun. Gene Cloning, Sequence Analysis and Expression Studies of Vitellogenin Gene in Geocoris pallidipennis[J]. Biotechnology Bulletin, 2015, 31(10): 149-156.

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