野生大豆GsGSK1基因的克隆及遗传转化

doi:10.3969/j.issn.1000-7091.2012.05.005

华北农学报 ›› 2012, Vol. 27 ›› Issue (5): 22-27. doi: 10.3969/j.issn.1000-7091.2012.05.005

所属专题： 油料作物；生物技术；

野生大豆GsGSK1基因的克隆及遗传转化

张春宝¹, 谭化¹, 赵丽梅¹, 彭浩², 吕龙石²

吉林省农业科学院大豆研究所;
延边大学农学院

收稿日期:2012-07-13 出版日期:2012-10-28
通讯作者: 赵丽梅(1964-)，女，吉林长春人，研究员，博士，主要从事大豆遗传育种研究。
作者简介:张春宝(1980-)，男，黑龙江佳木斯人，副研究员，博士，主要从事大豆分子生物学研究。
基金资助:
公益性行业(农业)科研专项(201003021);农业部野生资源保护与利用项目(2012);国家“863”计划项目(2011AA10A105)

Cloning and Genetic Transformation of Wild Soybean GsGSK1 Gene into Arabidopsis

ZHANG Chun-bao¹, TAN Hua¹, ZHAO Li-mei¹, PENG Hao², Lü Long-shi²

1. Soybean Research Institute Jilin Academy of Agricultural Sciences Changchun 130033 China;

2. College of Agriculture Yanbian University Yanji 133002 China

Received:2012-07-13 Published:2012-10-28

摘要/Abstract

摘要： 以野生大豆为材料,采用同源克隆方法和RT-PCR技术获得一个野生大豆糖原合成酶激酶类基因的全长cDNA(命名为GsGSK1)。该基因的ORF为1 233 bp,推断其编码410氨基酸的多肽。将此基因和与大豆糖原合成酶激酶基因的核苷酸序列比较发现二者有11个核苷酸SNP位点差异,其中,10个为同义突变,1个为非同义突变,同源性达到99.1%。通过构建重组植物表达载体pCAMBIA3300-GsGSK1,将该基因通过农杆菌介导法转入拟南芥,获得了T1抗性苗,为进一步鉴定GsGSK1的基因功能提供了试验基础。

关键词: 糖原合成酶激酶, 野生大豆, 转基因拟南芥, 抗逆性

Abstract: Using homologous cloning and RT-PCR technology,a glycogen synthase kinase family gene,GsGSK1 was isolated from wild soybean.It is 1 233 bp in length with one ORF of 410 amino acids.Compared the nucleotide sequence with GmGSK,these two genes have11 SNPs,in which 10 SNPs are synonymous mutations,and 1 SNP is non-synonymous mutation,with a homology of 99.1%.Then constructed the recombinant plant expression vector pCAMBIA3300-GsGSK1 and transformed it into arabidopsis by agrobacterium mediated method.T1 transgenic plant had been acquired,which will be supplied the basic materials for further identification of gene function in GsGSK1.

Key words: Glycogen synthase kinase, Wild soybean, Transgenic arabidopsis, Stress tolerance

中图分类号:

S565.1

张春宝, 谭化, 赵丽梅, 彭浩, 吕龙石. 野生大豆GsGSK1基因的克隆及遗传转化[J]. 华北农学报, 2012, 27(5): 22-27. doi: 10.3969/j.issn.1000-7091.2012.05.005.

ZHANG Chun-bao, TAN Hua, ZHAO Li-mei, PENG Hao, Lü Long-shi. Cloning and Genetic Transformation of Wild Soybean GsGSK1 Gene into Arabidopsis[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(5): 22-27. doi: 10.3969/j.issn.1000-7091.2012.05.005.

http://www.hbnxb.net/CN/Y2012/V27/I5/22

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