Molecular Cloning and Expression of cDNA Encoding the Cysteine Proteinase Inhibitor from Upland Cotton

被引:0
作者
Ming-feng Jiang
Sheng-wei Li
Min Chen
Ying-fan Cai
Yong-fang Xie
Biao Li
Quan Sun
Huai-zhong Jiang
Zheng Pan
Yun-ling Gao
You-Lu Yuan
Yu-zheng Shi
机构
[1] Southwest University for Nationalities,
[2] Chongqing University of Posts and Telecommunications,undefined
[3] Cotton Institute of Chinese Academy of Agricultural Science,undefined
来源
Journal of Plant Biology | 2009年 / 52卷
关键词
Cysteine proteinase inhibitor; cDNA cloning; Expression; RT-PCR; Western blotting;
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摘要
A cDNA encoding a novel cysteine proteinase inhibitor (CPI) was isolated from a gland mutant Xiangmian-18 of upland cotton during the pigments gland forming stage. The cDNA comprises 378 bp and encodes 125 amino acid residues with molecular mass of 13.8 kDa. It contains the conserved motif of cysteine protease inhibitors and belongs to the cystatin superfamily (Gln-Val-Val-Ala-Gly). The deduced amino acid sequences of the domains are highly similar to the normal upland cotton (96.8%). SDS-PAGE and western hybridization analysis showed that the expressed recombinant protein was recombinant CPI. The inhibitory activity of recombinant CPI was 46 u/μg which was measured by inhibiting the protease activity of papain. RT-PCR results indicated that the expression level of developing gland stage was higher than that of undeveloped gland stage.
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