烟青虫气味结合蛋白基因的克隆与序列分析

Molecular Cloning and Sequence Analysis of Genes Encoding GOBP1 and PBP in the Antenna of Helicoverpa assulta(Guenée)

根据已经报道的棉铃虫普通气味结合蛋白I(GOBP1)和性信息素结合蛋白(PBP)基因的cDNA序列,设计了2对引物,以烟青虫Helicoverpaassulta触角cDNA为模板,分别进行PCR扩增,获得2条特异性条带,约400bp。将以上两条片段分别连接到T-easy载体上,获得重组子T-GOBP1-Hass和T-PBP-Hass。序列测定和结构分析表明,Hass-GOBP1开放阅读框全长441bp,编码147个氨基酸残基,预测分子量和等电点分别为17.2kD和4.71。Hass-PBP开放阅读框全长405bp,编码135个氨基酸残基,预测分子量和等电点分别为15.1kD和5.2。Hass-GOBP1和Hass-PBP具有昆虫气味结合蛋白的典型特征,即氨基酸序列中具有6个保守的半胱氨酸残基,呈酸性。这2个基因已在GenBank中登记,序列号分别是AY864774和AY864775。

Genes encoding GOBP 1 and PBP in the antenna of Helicoverpa assulta （Guenée） were sequenced. Two pairs of primers were designed based on the comparison of GOBP1 and PBP gene sequences of Helicoverpa armigera reported previously. Two specific bands （about 400 bp in length） were amplified from cDNA of H. assulta antenna. Two segments were cloned into T-easy vector, respectively. Sequencing and structural analysis showed that the full-length of GOBP1 ORF was 441bp, 147 amino acid residues were encoded. The predicted MW and PI were 17.2 kD and 4.71, respectively. The full length of PBP ORF is 405bp, 135 amino acid residues were encoded. The predicted MW and PI were 15.1 kD and 5.2.The two sequences were deposited in GenBank and the accession number is AY8é4774 and AY864775.