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

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Abstract

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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Acknowledgments

This work was supported by grants from the National Nature Science Foundation of China (Nos. 30440032 and 30771311), Natural Sciences Foundation of Chongqing, China (Nos. cstc2007 BB1328), and the Committee of Sciences and Technology of Nan’an in Chongqing (2008), China.

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Authors and Affiliations

  1. Southwest University for Nationalities, Chengdu, 610041, China

    Ming-feng Jiang, Sheng-wei Li & Min Chen

  2. Chongqing University of Posts and Telecommunications, Chongqing, 400065, China

    Sheng-wei Li, Min Chen, Ying-fan Cai, Yong-fang Xie, Biao Li, Quan Sun, Huai-zhong Jiang, Zheng Pan & Yun-ling Gao

  3. Cotton Institute of Chinese Academy of Agricultural Science, Anyang, 455000, China

    You-Lu Yuan & Yu-zheng Shi

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  1. Ming-feng Jiang
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  2. Sheng-wei Li
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  3. Min Chen
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  5. Yong-fang Xie
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Correspondence to Ying-fan Cai.

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Ming-feng Jiang and Sheng-wei Li contributed equally to this work.

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Jiang, Mf., Li, Sw., Chen, M. et al. Molecular Cloning and Expression of cDNA Encoding the Cysteine Proteinase Inhibitor from Upland Cotton. J. Plant Biol. 52, 426–432 (2009). https://doi.org/10.1007/s12374-009-9054-6

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  • DOI: https://doi.org/10.1007/s12374-009-9054-6

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