[1]王羽,李红丽,王俊皓,等.大豆GmPM13基因的克隆及原核表达[J].大豆科学,2018,37(02):185-191.[doi:10.11861/j.issn.1000-9841.2018.02.0185]
　WANG Yu,LI Hong-li,WANG Jun-hao,et al.Cloning and Prokaryotic Expression of GmPM13 Gene from Glycine max[J].Soybean Science,2018,37(02):185-191.[doi:10.11861/j.issn.1000-9841.2018.02.0185]
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大豆GmPM13基因的克隆及原核表达

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第37卷 期数: 2018年02期 页码: 185-191 栏目: 出版日期: 2018-03-20
Title:
Cloning and Prokaryotic Expression of GmPM13 Gene from Glycine max
作者:
王羽; 李红丽; 王俊皓; 李海燕; 崔喜艳
（吉林农业大学 生命科学学院，吉林 长春 130118）
Author(s):
WANG Yu; LI Hong-li; WANG Jun-hao; LI Hai-yan; CUI Xi-yan
（School of Life Science，Jilin Agricultural University，Changchun 130118，China)
关键词:
大豆;  GmPM13基因;  基因克隆;  原核表达
Keywords:
Glycine max;  GmPM13 gene;  Gene cloning;  Prokaryotic expression
分类号:
S565.1；Q785
DOI:
10.11861/j.issn.1000-9841.2018.02.0185
文献标志码:
A
摘要:
通过对大豆（Glycine max）油体钙蛋白（caleosin）基因GmPM13的克隆及原核表达，为今后利用基因工程方法检测转GmPM13基因提供抗体。运用RT-PCR技术克隆得到大豆油体钙蛋白GmPM13基因，构建原核表达载体，命名为pET-28a-pm13。并转化到Ecdi和Rosetta中，经异丙基硫代半乳糖苷（IPTG）诱导后，SDS-PAGE分析GmPM13蛋白的表达情况。大豆GmPM13基因全长cDNA序列为738 bp，包括一个720 bp的开放阅读框，编码239个氨基酸，油体钙蛋白的分子量为27.1 kDa，诱导表达产物的大小与预计蛋白大小相符。在28℃，1.5 mol·L-1 IPTG浓度下，诱导12 h蛋白的表达量最高，占总蛋白的39.25％。
Abstract:
Through the cloning and prokaryotic expression of caleosin gene GmPM13 in soybean could provid the antibody for the detection of GmPM13 gene by genetic engineering in the future. GmPM13 gene of soybean caleosin was cloned by RT-PCR technology, and the prokaryotic expression vector was constructed which was named GmPM13. It was transformed into E.coli Rosetta with IPTG induction, the expression of GmPM13 protein was analyzed by SDS-PAGE. The full-length cDNA sequence of GmPM13 gene was 738 bp with a 720 bp ORF(open reading frame), encoding 239 amino acids. The molecular mass of the protein was 27.1 kDa, and the size of the induced product was consistent with the expected protein size. The induced protein content reached the highest level at 28℃, 1.5 mol·L-1 IPTG concentration, after induced 12 h, accounting for 39.25% in the total bacterial proteins.

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备注/Memo

收稿日期：2017-10-19

基金项目：国家自然科学基金(31171459)；吉林省教育厅“十三五”科学技术项目基金(JJKH20170307KJ)；吉林省自然科学基金(20180101026JC)。
第一作者简介：王羽（1993-），女，硕士，主要从事生物化学与分子生物学研究。E-mail:448659230@qq.com。
通讯作者：崔喜艳（1971-），女，博士，教授，主要从事生物化学与分子生物学研究。E-mail:cuixiyan2005@163.com。

更新日期/Last Update: 2018-04-02