红脊长蝽感觉神经元膜蛋白基因克隆及组织表达谱分析

Cloning and Expression Pattern of Sensory Neuron Membrane Protein Genes of Tropidothorax elegans

[目的]感觉神经元膜蛋白(SNMP)是至关重要的次要嗅觉蛋白,能够作为信息素识别的标记,而红脊长蝽(Tropidothorax elegans)作为一个重要的农林害虫,这方面的研究还很少。[方法]利用PCR方法对红脊长蝽SNMP基因进行克隆,并通过荧光定量PCR对该基因的表达情况进行分析。[结果]首次克隆和鉴定了红脊长蝽的2个SNMPs基因,并命名为TeleSNMP1和TeleSNMP2。序列分析表明,TeleSNMP1编码区开放阅读框长1497 bp,编码498 aa;而TeleSNMP2编码区开放阅读框长1686 bp,编码561 aa,这2个基因的等电点分别为8.26和7.05。同源性分析发现,同目昆虫同类SNMP序列一致性较高,不同目昆虫不同类SNMP序列一致性较低。TeleSNMP1和TeleSNMP2之间的序列一致性极低。进化树结果也表现同目昆虫同类SNMP基因进化关系最近。定量PCR结果显示,TeleSNMP1主要在雌雄触角中表达,而TeleSNMP2在非触角组织中也有表达。[结论]该结果为今后对红脊长蝽SNMPs基因功能的研究提供了有用的参考。

[Objective]To study the sensory neuron membrane proteins(SNMPs)in Tropidothorax elegans.[Method]The SNMP genes were cloned by PCR,and the expression of these genes were analyzed by fluorescence quantitative PCR.[Result]Two SNMPs genes of T.elegans,named as TeleSNMP1 and TeleSNMP2,were cloned and identified for the first time by RT-PCR.Sequence analysis showed that the TeleSNMP1 encoding area was 1497 bp in length,encoding 498 aa.The TeleSNMP2 encoding area was 1686 bp in length,encoding 561 aa.The isoelectric points of these two genes were 8.26 and 7.05,respectively.Homology analysis showed that the consistency of SNMPs genes in insects of the same order was high,while the consistency of different order was low,and the sequence consistency between TeleSNMP1 and TeleSNMP2 was extremely low.The evolutionary tree results also showed that the evolutionary relationship of SNMPs genes in insects of the same order was rather close.Quantitative real-time PCR showed that TeleSNMP1 was antennae-biased,whereas TeleSNMP2 was expressed in female and male antennae but was also expressed in nonantennal tissues.[Conclusion]The results provide a useful reference for future studies on the function of the T.elegans SNMPs gene.