松墨天牛寡糖基转移酶亚基<em>STT3B</em>的克隆与表达分析

doi:10.7668/hbnxb.2016.03.012

华北农学报 ›› 2016, Vol. 31 ›› Issue (3): 80-87. doi: 10.7668/hbnxb.2016.03.012

所属专题： 畜牧；

松墨天牛寡糖基转移酶亚基STT3B的克隆与表达分析

蔡紫玲, 吴华俊, 林同

华南农业大学林学与风景园林学院, 广东广州 510642

收稿日期:2016-03-12 出版日期:2016-06-28
通讯作者: 林同(1969-),男,黑龙江通河人,副教授,博士,主要从事昆虫分子生物学研究。
作者简介:蔡紫玲(1992-),女,广东茂名人,在读硕士,主要从事昆虫分子生物学研究。
基金资助:
国家自然科学基金项目(31470653);广东省自然科学基金项目(1414050001666)

Cloning and Expression Analysis of STT3B of Oligosaccharyltransferase from Monochamus alternatus

CAI Ziling, WU Huajun, LIN Tong

College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou 510642, China

Received:2016-03-12 Published:2016-06-28

摘要/Abstract

摘要： 为阐明松墨天牛寡糖基转移酶亚基STT3B的结构特点和在不同虫态、成虫的不同部位以及幼虫组织中的表达情况,采用反转录聚合酶链式反应(RT-PCR)和快速扩增cDNA末端(RACE)技术克隆松墨天牛STT3B基因cDNA,命名为MaSTT3B,其GenBank登录号为KT362368。序列分析显示其长度为2552bp,其中开放阅读框长2322bp,编码773个氨基酸。MaSTT3B的氨基酸序列与赤拟谷盗相似性最高,为93%,在系统发育树的同一个分支上。用实时荧光定量PCR分析MaSTT3B的相对表达量,结果表明:STT3B在蛹中的表达量高于成虫;在成虫足的表达量最高;在幼虫各组织中,体壁的相对表达量最高。

关键词: 松墨天牛, STT3B, 基因克隆, RT-qPCR

Abstract: In order to analyze the structure characteristics of MaSTT3B and its expression in the different development stages,different parts of adults and different tissues in larvae,we cloned the STT3B gene of Monochamus alternatus named MaSTT3B (GenBank accession number:KT362368) using the technology including reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification (RACE).The total sequence length of MaSTT3B is 2 552 bp,including 2 322 bp open reading frame encoding 773 amino acids.The amino acid sequence of MaSTT3B has 93% similarity to Tribolium castaneum, and both of them are in the same branch of phylogenetic tree based on insect STT3B.The result of RT-qPCR indicated that MaSTT3B was expressed higher in pupae than in adults,in adult's legs than in other parts of adults,and highest in body wall of among larvae tissues.The relative expression levels of MaSTT3B was detected by RT-qPCR indicated that MaSTT3B was expressed higher in pupae than in adults,in adult's legs than in other parts of adults,and highest in body wall of among larvae tissues.

Key words: Monochamus alternatus, STT3B, Gene cloning, RT-qPCR

中图分类号:

Q966
Q78

蔡紫玲, 吴华俊, 林同. 松墨天牛寡糖基转移酶亚基STT3B的克隆与表达分析[J]. 华北农学报, 2016, 31(3): 80-87. doi: 10.7668/hbnxb.2016.03.012.

CAI Ziling, WU Huajun, LIN Tong. Cloning and Expression Analysis of STT3B of Oligosaccharyltransferase from Monochamus alternatus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(3): 80-87. doi: 10.7668/hbnxb.2016.03.012.

http://www.hbnxb.net/CN/Y2016/V31/I3/80

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松墨天牛寡糖基转移酶亚基STT3B的克隆与表达分析

Cloning and Expression Analysis of STT3B of Oligosaccharyltransferase from Monochamus alternatus

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松墨天牛寡糖基转移酶亚基STT3B的克隆与表达分析

Cloning and Expression Analysis of STT3B of Oligosaccharyltransferase from Monochamus alternatus

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