松墨天牛原肌球蛋白基因的克隆和表达检测

利用先前构建的cDNA文库和RACE方法,克隆了松墨天牛原肌球蛋白基因的cDNA全长,该序列长1 203 bp,命名为MaTm(GenBank登录号:KM099072),其中开放阅读框长852 bp,编码283个氨基酸.MaTm的氨基酸序列与赤拟谷盗 (Tribolium castaneum)相似性最高,为97%,与家蚕相似性 (Bombyx mori)为94%.松墨天牛与赤拟谷盗处在系统发育树的同一个分支上.用 RT-qPCR 分析了MaTm的相对表达量,结果显示:蛹和幼虫的表达量低于成虫;成虫足和头的表达量高于对照;在幼虫各组织中均有表达,且差异显著(p0.05),其中在体壁的表达量最高.

The cloning and expression of tropomyosin gene from Monochamus alternatus

The cDNA of tropomyosin gene from Monochamus alternatus with the full length of 1133 bp,designated as MaTm (GenBank:KM099072),was cloned by using the cDNA library previously constructed and rapid-amplification of cDNA ends (RACE). MaTm contains an ORF (open reading frame)which is comprised of 852 bp,encoding 283 amino acid residues.MaTm showed 97% identity with Tribolium castaneum,94% identity with Bombyx mori,and on the same branch in the phylogenetic tree based on insect tropomyosin amino acids.The expression characteristics analyzed by RT-qPCR indicated that the expression levels of MaTm in pupae and larvae were lower than that in adults,and the expression in head and legs of adults were higher than that in control.MaTm was prominent differently expressed in all of the detected larval tissues(p0.05),with the highest expression level in body wall.