|
|||||
|
|
| sMiMi-CK cDNA的克隆、表达及活性检测 | |
| 引用本文： | 李芳,李守岩,王孝铭,关振中. sMiMi-CK cDNA的克隆、表达及活性检测[J]. 中国急救医学, 1999, 19(5): C19 |
| 作者姓名： | 李芳 李守岩 王孝铭 关振中 |
| 作者单位： | 1. 哈尔滨医科大学病生教研室,150086 2. 黑龙江省前哨农场医院 3. 哈医大附属第二医院心内科 |
| 摘 要： | 目的本实验拟通过ＲＴ－ＰＣＲ技术，克隆及表达大鼠心肌线粒体ｓＭｉＭｉ－ＣＫ基因。方法从大鼠心肌线粒体总ＲＮＡ中扩增出ｓＭｉＭｉ－ＣＫＤＮＡ片段（１３３ｋｂ），将该片段克隆到载体ｐＵＣ１９上，对其进行酶切和测序，然后将其克隆到ＰＢＶ２２０上，经温度诱导，使其在大肠杆菌中得以表达，并用ＮＡＤＨ法测其活性。结果ｓＭｉＭｉ－ＣＫ基因序列与国外报导的序列完全一致，表达的ｓＭｉＭｉ－ＣＫ分子量为４５８ＫＤ，占菌体总蛋白的１０７％，其活性为２７４×１０５ＩＵ／Ｌ菌液。结论ｓＭｉＭｉ－ＣＫ基因的克隆及表达，为今后从基因水平上研究心肌缺血性损伤的机制及治疗提供了依据。 |
| 关 键 词： | sMiMi－CK RT－PCR 克隆 表达 |
Clone, expression and activity of rat myocardial mitochondrial sarcomeric creaine kinase |
|
| Li Fang,Li Shouyan,Wang Xiaoming,et al.. Clone, expression and activity of rat myocardial mitochondrial sarcomeric creaine kinase[J]. Chinese Journal of Critical Care Medicine, 1999, 19(5): C19 | |
| Authors: | Li Fang Li Shouyan Wang Xiaoming et al. |
| Affiliation: | Li Fang,Li Shouyan,Wang Xiaoming,et al.Departmet of pathophysilogy,Harbin Medical University,Harbin 150086 |
| Abstract: | Objective A DNA fregment which code rat myocardial mitochondrial sarcomeric creatine kinase (sMiMi-CK) has been amplified by RT-PCR from rat myocardial genomic RNA.It was colned and expressed. Methods sMiMi-CK gene was cloned to plasmid pUC19 and subjected to sequenced analysis.An expression clone of sMiMi-CK was constructed PBV 220,and protein activity can be measured by automatic bitchem. Results The DNA sequenceis the same as reported abroad.The recombinated sMiMi-CK was stenthesized in an Escherichia coli expression system and its molecular weight was about 45.8KD,the expressed sMiMi-CK was about 10.7 per cent of total bacteria protein.Its activity was 2.74105 IU/L in colibacillus. Conclusion sMiMi-CK colne and expression provide some useful data for the study of cardiac ischemia injury mechanism and treatmeat in the gene levels. |
| Keywords: | MiMi-CKRT-PCRCloneExpression |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |