| 厚壳贻贝M7 lysin分子的克隆与表达 |
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| 引用本文： | 刘慧慧,薛超波,常抗美,杨刚.厚壳贻贝M7 lysin分子的克隆与表达[J].水产学报,2011,35(9):1337-1342. |
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| 作者姓名： | 刘慧慧 薛超波 常抗美 杨刚 |
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| 作者单位： | 1. 浙江海洋学院海洋科学学院,浙江省海洋养殖装备与工程技术重点实验室,浙江舟山316000
2. 舟山市质量技术监督检测院,浙江舟山,316000 |
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| 基金项目： | 浙江省自然科学基金资助项目(Y307381);国家科技支撑计划(2011BAD13B08);“海洋渔业科学与技术”省重中之重学科开放课题(20100202) |
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| 摘 要： | M7 lysin位于贻贝精子顶体中,是溶解卵黄膜、促进受精作用的重要蛋白质,决定了贻贝种间精卵识别的特异性。采用同源克隆法得到厚壳贻贝M7 lysin分子,并在原核生物中对该分子进行重组表达。结果表明,扩增产物为540 bp左右的片段,进一步测序发现其开放阅读框cDNA为543 bp,与贻贝、地中海贻贝、盖勒贻贝具有较高的相似性;在线翻译所得蛋白质片段中含有180个氨基酸,分子量为20 ku,等电点为8.48;通过M7 lysin氨基酸序列构建系统进化树发现,贻贝和地中海贻贝亲缘关系最近,其次为盖勒贻贝,最后是厚壳贻贝;将 M7 lysin分子在E.coli Rosseta(DE3)中融合表达,得到包括载体氨基酸序列在内的25 ku蛋白质,符合预期分子质量。
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| 关 键 词： | 厚壳贻贝 精子 M7 lysin 表达 |
| 收稿时间： | 2011/3/26 0:00:00 |
| 修稿时间： | 5/26/2011 3:33:43 PM |
The cloning and expression of M7 lysin in Mytilus coruscus |
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| LIU Hui-hui,XUE Chao-bo,CHANG Kang-mei and YANG Gang.The cloning and expression of M7 lysin in Mytilus coruscus[J].Journal of Fisheries of China,2011,35(9):1337-1342. |
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| Authors: | LIU Hui-hui XUE Chao-bo CHANG Kang-mei and YANG Gang |
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| Institution: | Marine Science School of Zhejiang Ocean University,Key Laboratory of Zhejiang Provincial Marine Culturing Equipments and Engineering Technology,Zhoushan Center of Calibration and Testing for Qualitative Technical Supervision,Marine Science School of Zhejiang Ocean University,Key Laboratory of Zhejiang Provincial Marine Culturing Equipments and Engineering Technology,Marine Science School of Zhejiang Ocean University,Key Laboratory of Zhejiang Provincial Marine Culturing Equipments and Engineering Technology |
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| Abstract: | M7 lysin is located in the mussel sperm acrosome.As an important protein for fertilization,it can dissolve vitelline membrane and determine the specificity of sperm-egg recognition of mussel.At present,the M7 lysin sequences of Mytilus edulis Linnaeus,M.galloprovincialis and M.trossulus have been recognized,but it hasn't been reported in M.coruscus.In this study,we cloned M7 lysin of M.coruscus with homology cloning method,and it was expressed in E.coli Rosseta(DE3).The results showed that the amplified pro... |
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| Keywords: | Mytilus coruscus sperm M7 lysin expression |
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