[1]李培培,张皛,潘小玫,等.截短的小麦冷胁迫反应蛋白基因片段TA3-13的融合表达、纯化与诱导抗TMV功能研究[J].南京农业大学学报,2010,33(1):54-58.[doi:10.7685/j.issn.1000-2030.2010.01.011]
 LI Pei-pei,ZHANG Xiao,PAN Xiao-mei,et al.His-tag fusion expression,purification and TMV resistance induction of truncated wheat cold shock protein gene fragment TA3-13[J].Journal of Nanjing Agricultural University,2010,33(1):54-58.[doi:10.7685/j.issn.1000-2030.2010.01.011]
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截短的小麦冷胁迫反应蛋白基因片段TA3-13的融合表达、纯化与诱导抗TMV功能研究()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
33卷
期数:
2010年1期
页码:
54-58
栏目:
出版日期:
2010-01-31

文章信息/Info

Title:
His-tag fusion expression,purification and TMV resistance induction of truncated wheat cold shock protein gene fragment TA3-13
作者:
李培培张皛潘小玫王金生宋从凤
南京农业大学植物保护学院/农业部病虫监测与治理重点开放实验室,江苏,南京,210095
Author(s):
LI Pei-pei ZHANG Xiao PAN Xiao-mei WANG Jin-sheng SONG Cong-feng
College of Plant Protection/Key Laboratory of Monitoring and Management of Plant Diseases and Insects, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
关键词:
小麦 TA3-13 His融合表达 纯化 TMV抗性
Keywords:
wheat TA3-13 His-tag fusion expression purification TMV resistance
分类号:
S512.1
DOI:
10.7685/j.issn.1000-2030.2010.01.011
文献标志码:
A
摘要:
TA3-13是小麦中一个冷胁迫蛋白WCSP1编码基因的5′端截短的DNA序列。它与水稻白叶枯病菌Harpin蛋白的编码基因hpa1_{X∞}(又称hrf1,hrfA)具有59%的一致性。为了研究TA3-13蛋白的生物学性状和功能及其与Harpin蛋白的关系,本研究以pET30a(+)为载体,构建了His融合表达重组体,转化大肠杆菌后在异丙基硫代-β-D-半乳糖苷(IPTG)和阿拉伯糖的诱导下获得了融合蛋白的表达。利用镍柱对融合蛋白进行了纯化研究,结果表明:融合蛋白在200 mmol·L^{-1}咪唑浓度下能够被有效地洗脱下来,电泳显示具有单一条带。烟草花叶病毒(TMV)接种试验显示:纯化的TA3-13蛋白喷雾烟草后仍具有较高的诱导烟草抗TMV的作用。
Abstract:
TA3-13 is a 5’truncated sequence of a wheat gene that encodes the cold shock protein WCSP1.TA3-13 shares 59% identity with hpa1_{Xoo}, a Harpin encoding gene of Xanthomonas oryzae pv. oryzae. To study the biological function of TA3-13, we made a His-tag fusion expression construct that could express the fusion protein under the induction of isopropylthio-beta-D-galactopyranoside (IPTG) and arabinose. The fusion protein was purified by nickel chromatograph and eluted with 200 mmol·L^{-1} imidazole. When applied to tobacco by spraying plant leaf, the purified TA3-13 protein induced resistance to tobacco mosaic virus.

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备注/Memo

备注/Memo:
国家自然科学基金项目(30671355);国家973计划项目(2003CB114204);江苏省科技攻关项目(BE-2005-604)
更新日期/Last Update: 2010-01-31