[1]王丕武　刘灵芝　柴晓杰　张君　曲同宝.大豆11S 球蛋白Gy5(A3B4)cDNA 的克隆及植物表达载体的构建[J].大豆科学,2004,23(03):159-163.[doi:10.11861/j.issn.1000-9841.2004.03.0159]
　Wang Peiw u,Liu Lingzhi,Chai Xiaojie,et al.cDNA CLONING AND PLANT EXPRESSION VECTOR CONSTRUCTIONOF SOYBEAN 11S GLYCUININ Gy5[J].Soybean Science,2004,23(03):159-163.[doi:10.11861/j.issn.1000-9841.2004.03.0159]
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大豆11S 球蛋白Gy5(A3B4)cDNA 的克隆及植物表达载体的构建

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第23卷 期数: 2004年03期 页码: 159-163 栏目: 出版日期: 2004-08-25
Title:
cDNA CLONING AND PLANT EXPRESSION VECTOR CONSTRUCTIONOF SOYBEAN 11S GLYCUININ Gy5
文章编号:
1000 -9841(2004)03 -0159 -05
作者:
王丕武　刘灵芝　柴晓杰　张君　曲同宝
吉林农业大学生物中心, 长春130118
Author(s):
Wang Peiw u ;  Liu Lingzhi ;  Chai Xiaojie ;  Zhang Jun ;  Qu Tongbao
Biotechnology Center of Ji lin Agricultural University , Changchun 130118)
关键词:
大豆; Gy 5 ; cDNA ; 克隆; 植物表达载体
Keywords:
Soybean ; Gy 5 ; cDNA ; Clone ; Plant expression vector
分类号:
S 565.1
DOI:
10.11861/j.issn.1000-9841.2004.03.0159
文献标志码:
A
摘要:
从大豆(花生豆1 号)未成熟种子中提取总RNA , 逆转录形成cDNA 第一条链。根据genebank 中大豆11S 球蛋白Gy5 的cDNA 序列设计引物, 用PCR 方法扩增Gy5 的cDNA 序列, 克隆到pUC19 质粒载体上, 并测定其全序列。用限制性内切酶PstI 和EcoRI 酶切重组质粒, 将目的片段与质粒pCAMBIA1301 连接, 构建成植物表述载体并转化到农杆菌中, 以便于以后的研究利用。
Abstract:
The total RNA w as est racted f rom immature seeds of soybean and direct ly used in single -stranded(ss)DNA synthetic reaction .According to soybean 11S g ly cinin Gy5 cDNA sequences in genebank , a pair ofprimers were designed .Double -stranded(ds)cDNA w as amplified by PCR 。The amplif ied Gy5 cDNA wascloned into plasmid pUC 19 and sequenced 。Fragment of the Gy5 cDNA in the recombinant plasmid w as cutw ith Pst I/EcoRI and ligated wi th plasmid Pcambia1301 , then a plant expression vector w as const ructed 。It wasalso t ransferred into Agrobacterium LBA4404 in o rder to be utilized later.

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备注/Memo

基金项目:“ 国家转基因植物中试与产业化基地(吉林)” 专项课题(J99 -B -001)　

作者简介:王丕武(1958 -), 男, 教授, 主要从事大豆遗传育种研究。

更新日期/Last Update: 2017-01-01