烟夜蛾感觉神经元膜蛋白基因的克隆、序列分析与组织特异性表达

Cloning,sequence analysis and tissue-specific expression of sensory neuron membrane protein gene from Helicoverpa assulta(Guenée)

利用反转录多聚酶链式反应(RT-PCR)技术从烟夜蛾Helicoverpa assulta(Guenée)触角中扩增得到感觉神经元膜蛋白(sensory neuron membrane protein,SNMP)基因HassSNMP(GenBank登录号:EU580138)。序列分析结果显示,该片段长度为1658bp,其中ORF长1572bp,编码523个氨基酸残基,预测成熟蛋白分子量为59.2 kDa,等电点为6.54。序列比对结果表明,烟夜蛾与其它昆虫的SNMP基因推导表达的氨基酸序列具有较高的一致性。半定量RT-PCR研究结果显示,SNMP在烟夜蛾触角、头(去掉触角)、喙和足中表达,其中在触角中的表达量最高,且雌雄虫触角中的表达量差异不显著,在喙中的表达量次之,在翅、胸和腹中没有表达;此外,该基因在成虫、十日龄蛹以及卵中也有表达,其中在卵中的表达量相对较低;在幼虫、一日龄蛹、四日龄蛹和七日龄蛹中没有表达。该研究结果说明烟夜蛾SNMP是非触角特异性表达基因。

Abstract： The gene HassSNMP （registered with GenBank accession no. EU580138） encoding sensory neuron mem- brane protein （SNMP） from antennae of Helicoverpa assulta （Guen6e） was cloned by reverse transcription polymerase chain reaction （RT-PCR）. Results of cloning and sequencing showed that the full-length open reading-frame （ORF） of SNMP is 1572 bp encoding 523 amino acid residues. The predicted molecular weight （MW） and isoelectric point （pI） are 59. 2 kDa and 6.54, respectively. The deduced amino acids were further aligned with sequences of SNMPs from oth- er insects. This results indicated that the predicted protein obtained shares high identity with other known SNMPs. The gene was hence named as HassSNMP. Additionally, semi-quantitative RT-PCR analysis showed that HassSNMP is not only expressed in antennae, but also in heads （antennae excluded）, proboscis and legs of H. assulta. The most robust expression was observed in antennae. Similar expression patterns were observed separately in both male and female an- tennae. Moreover, HassSNMP was highly expressed in proboscises but not in wings, thoraxes and abdomens. HassSNMP was also expressed in andult, 10th-day pupae and embryos, of which low expressed in embryos. However, HassSNMPwas not expressed in larvae, 1 st-day pupae, 4th-day pupae and 7th-day pupae. Altogether, these results suggested that HassSNMP is non-antenna specific gene.