Molecular Cloning, Recombinant Expression and Antifungal Activity of BnCPI, a Cystatin in Ramie (Boehmeria nivea L.)

文献类型： 外文期刊

第一作者： Yu, Yongting

作者： Yu, Yongting;Li, Zhimin;Cheng, Yi;Gao, Chunsheng;Zeng, Liangbin;Chen, Jia;Yan, Li;Sun, Xiangping;Guo, Litao;Yan, Zhun;Yu, Yongting;Li, Zhimin;Cheng, Yi;Gao, Chunsheng;Zeng, Liangbin;Chen, Jia;Yan, Li;Sun, Xiangping;Guo, Litao;Yan, Zhun;Zhang, Gang;Zhang, Gang

作者机构：

关键词： Boehmeria nivea L.;BnCPI;cysteine protease inhibitors;intron;functional expression

期刊名称：GENES （ 影响因子：4.096； 五年影响因子：4.339 ）

ISSN： 2073-4425

年卷期： 2017 年 8 卷 10 期

页码：

收录情况： SCI

摘要： Phytocystatins play multiple roles in plant growth, development and resistance to pests and other environmental stresses. A ramie (Boehmeria nivea L.) phytocystatin gene, designated as BnCPI, was isolated from a ramie cDNA library and its full-length cDNA was obtained by rapid amplification of cDNA ends (RACE). The full-length cDNA sequence (691 bp) consisted of a 303 bp open reading frame (ORF) encoding a protein of 100 amino acids with deduced molecular mass of 11.06 kDa and a theoretical isoelectric point (pI) of 6.0. The alignment of genome DNA (accession No. MF153097) and cDNA sequences of BnCPI showed that an intron (similar to 104 bp) exists in the coding region. The BnCPI protein contains most of the highly conserved blocks including Gly(5)-Gly(6) at the N-terminal, the reactive site motif QxVxG (Q(49)V(50)V(51)S(52)G(53)), the L-79-W-80 block and the [LVI]-[AGT]-[RKE]-[FY]-[AS]-[VI]-x-[EDQV]-[HYFQ]-N (L(22)G(23)R(24) F(25)A(26)V(27) (DDH30)-D-28-H-29 N-31) block that is common among plant cystatins. BLAST analysis indicated that BnCPI is similar to cystatins from Glycine max (77%), Glycine soja (76%), Hevea brasiliensis (75%) and Ricinus communis (75%). The BnCPI was subcloned into expression vector pSmart-I and then overexpressed in Escherichia coli BL21 (DE3) as a His-tagged recombinant protein. The purified reBnCPI has a molecular mass of 11.4 kDa determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Purified reBnCPI can efficiently inhibit the protease activity of papain and ficin toward BANA (N alpha-benzoyl-L-arginine-2-naphthyamide), as well as the mycelium growth of some important plant pathogenic fungi. The data further contribute to our understanding of the molecular functions of BnCPI.

分类号：

  • 相关文献

[1]Cloning and Characterization of a GABA Receptor from Plutella xylostella (Lepidoptera: Plutellidae). Zhou, Xiao-Mao,Wu, Qing-Jun,Zhou, Xiao-Mao,Zhang, You-Jun,Bai, Lian-Yang,Huang, Xiong-Ying.

[2]Functional expression of the keratinolytic serine protease gene sfp2 from Streptomyces fradiae var. k11 in Pichia pastoris. Li, Jiang,Shi, Peng-Jun,Han, Xiao-Yu,Meng, Kun,Yang, Pei-Long,Wang, Ya-Ru,Luo, Hui-Ying,Wu, Ning-Feng,Yao, Bin,Fan, Yun-Liu.

[3]Expression of five AtHsp90 genes in Saccharomyces cerevisiae reveals functional differences of AtHsp90s under abiotic stresses. Song, Hongmiao,Fan, Pengxiang,Shi, Wuliang,Li, Yinxin,Song, Hongmiao,Zhao, Rongmin. 2010

[4]Sequence variation of Bemisia tabaci Chemosensory Protein 2 in cryptic species B and Q: New DNA markers for whitefly recognition. Liu, Guo-xia,Ma, Hong-mei,Xie, Hong-yan,Xuan, Ning,Picimbon, Jean-Francois,Picimbon, Jean-Francois,Picimbon, Jean-Francois. 2016

[5]Mitochondrial genomes of the green macroalga Ulva pertusa (Ulvophyceae, Chlorophyta): novel insights into the evolution of mitogenomes in the Ulvophyceae. Liu, Feng,Liu, Feng,Melton, James T., III,Bi, Yuping. 2017

[6]Cloning and Sequence Analysis of Prophenoloxidase from Haemocytes of the Red Swamp Crayfish, Procambarus clarkii. Li Yan-he,Chen Hong-quan,Zheng Fang-liang,Wang Han-zhong,Li Yan-he,Wang Liu-quan,Xu Di-ping. 2009

[7]The Long Intron 1 of Growth Hormone Gene from Reeves' Turtle (Chinemys reevesii) Correlates with Negatively Regulated GH Expression in Four Cell Lines. Liu, Wen-Sheng,Ma, Jing-E,Li, Wei-Xia,Zhang, Jin-Ge,Wang, Juan,Nie, Qing-Hua,Qiu, Feng-Fang,Fang, Mei-Xia,Zeng, Fang,Wang, Xing,Lin, Xi-Ran,Zhang, Li,Chen, Shao-Hao,Zhang, Xi-Quan,Liu, Wen-Sheng,Ma, Jing-E,Li, Wei-Xia,Zhang, Jin-Ge,Wang, Juan,Nie, Qing-Hua,Qiu, Feng-Fang,Fang, Mei-Xia,Zeng, Fang,Wang, Xing,Lin, Xi-Ran,Zhang, Li,Chen, Shao-Hao,Zhang, Xi-Quan,Liu, Wen-Sheng,Ma, Jing-E,Li, Wei-Xia,Zhang, Jin-Ge,Wang, Juan,Nie, Qing-Hua,Qiu, Feng-Fang,Fang, Mei-Xia,Zeng, Fang,Wang, Xing,Lin, Xi-Ran,Zhang, Li,Chen, Shao-Hao,Zhang, Xi-Quan,Liu, Wen-Sheng. 2016

[8]Functional analysis of the larval serum protein gene promoter from silkworm, Bombyx mori.. Tang, SM,Yi, YZ,Shen, XJ,Zhang, ZF,Li, YR,He, JL. 2003

[9]Inferring Invasion History of Red Swamp Crayfish ( Procambarus clarkii) in China from Mitochondrial Control Region and Nuclear Intron Sequences. Li, Yanhe,Chen, Liping,Bai, Xiaohui,Wei, Xinlan,Zhou, Xiaoyun,Huang, Songqian,Wang, Weimin,Li, Yanhe,Guo, Xianwu,Bai, Xiaohui. 2015

[10]Rapid Construction of Stable Infectious Full-Length cDNA Clone of Papaya Leaf Distortion Mosaic Virus Using In-Fusion Cloning. Tuo, Decai,Shen, Wentao,Yan, Pu,Li, Xiaoying,Zhou, Peng. 2015

[11]Characterization of the promoter and 5 '-UTR intron of oleic acid desaturase (FAD2) gene in Brassica.napus. Xiao, Gang,Lu, Chang Ming,Xiao, Gang,Tan, Tai Long,Guan, Chun Yun,Xiao, Gang,Zhang, Zhen Qian,Yin, Chang Fa,Liu, Rui Yang,Wu, Xian Meng,Tan, Tai Long,Chen, She Yuan,Guan, Chun Yun. 2014

[12]Functional Analysis of the ZAG2 Promoter from Maize in Transgenic Tobaccos. Lu Min-hui,Wang Jian-hua,Wang Guo-ying,Meng Zheng. 2012

[13]Molecular Characterization of High Plant Species Using PCR with Primers Designed from Consensus Branch Point Signal Sequences. Xiong, Faqian,Jiang, Jing,Han, Zhuqiang,Zhong, Ruichun,He, Liangqiong,Tang, Ronghua,Zhuang, Weijian. 2011

[14]An AGAMOUS intron-driven cytotoxin leads to flowerless tobacco and produces no detrimental effects on vegetative growth of either tobacco or poplar. Li, Wei,Hu, Wei,Fang, Chu,Chen, Longzheng,Zhuang, Weibing,Katin-Grazzini, Lorenzo,McAvoy, Richard J.,Guillard, Karl,Li, Yi,Chen, Longzheng,Zhuang, Weibing,Li, Yi,Zhuang, Weibing,Li, Yi.

作者其他论文 更多>>

微信小程序