柠条锦鸡儿<em>CkLEA4-1</em>基因的克隆及表达分析

doi:10.7668/hbnxb.2017.02.015

摘要/Abstract

摘要： 为研究胚胎发育晚期蛋白（LEA）在柠条中的生物学功能，从已构建的柠条锦鸡儿干旱胁迫抑制削减杂交文库中筛选到一条LEA蛋白编码基因，并采用RT-PCR法进行克隆。所得LEA基因的开放阅读框（ORF）长1 119 bp，编码373个氨基酸的蛋白质，命名为CkLEA4-1。结合序列比对与系统进化分析结果，推断CkLEA4-1属于5族LEA蛋白。利用实时荧光定量PCR技术对CkLEA4-1在干旱、高盐等逆境胁迫条件下的表达情况进行初步研究，结果表明，CkLEA4-1受到不同程度的诱导，推测该基因可能与柠条锦鸡儿响应逆境胁迫有关。研究还成功构建了CkLEA4-1的过表达载体pCanG-CkLEA4-1，得到转基因纯合体株系，为进一步研究柠条锦鸡儿CkLEA4-1基因的功能提供了材料。

关键词: 柠条锦鸡儿, LEA, 基因克隆, 表达分析

Abstract: In order to research the biological function of the LEA protein,a gene encoding LEA protein from the suppression subtractive hybridization library of Caragana korshinskii Kom.was screened and cloned by using RT-PCR method.The open reading frame (ORF)of the gene length was 1 119 bp,encoded a protein of 373 amino acids,named CkLEA4-1.Based on sequence alignment and phylogenetic analysis,it was inferred that CkLEA4-1 belonged to group 5 late embryogenesis abundant protein (LEA).The expression of CkLEA4-1 under high salt and drought conditions were detected by Real-time quantitative PCR,the results showed that CkLEA4-1 could be induced at different levels,which suggested that the gene might be involved in stress response of Caragana korshinskii Kom..The over-expression vector pCanG-CkLEA4-1 of CkLEA4-1 was constructed successfully,and the transgenic homozygous lines were obtained,which provided materials for the further study of CkLEA4-1 gene function in Caragana korshinskii Kom.

Key words: Caragana korshinskii Kom., LEA, Gene cloning, Expression analysis

中图分类号:

S793.3

滑璐玢, 于秀敏, 杨杞, 王瑞刚, 宝力德. 柠条锦鸡儿CkLEA4-1基因的克隆及表达分析[J]. 华北农学报, 2017, 32(2): 96-103. doi: 10.7668/hbnxb.2017.02.015.

HUA Lubin, YU Xiumin, YANG Qi, WANG Ruigang, BAO Lide. Clone and Expression Analysis of CkLEA4-1 from Caragana korshinskii Kom.[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(2): 96-103. doi: 10.7668/hbnxb.2017.02.015.

http://www.hbnxb.net/CN/Y2017/V32/I2/96

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柠条锦鸡儿CkLEA4-1基因的克隆及表达分析

Clone and Expression Analysis of CkLEA4-1 from Caragana korshinskii Kom.

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柠条锦鸡儿CkLEA4-1基因的克隆及表达分析

Clone and Expression Analysis of CkLEA4-1 from Caragana korshinskii Kom.

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