中文摘要：

目的本研究旨在建立心肌特异性表达人KCNE4（hKcNE4）基因小鼠模型，为探索hKCNE4基因对特定类型心房颤动的防治作用奠定基础。方法通过RT—PCR克隆hKCNE4基因，使用核酸内切酶和连接酶构建hKCNE4基因心肌特异性表达载体hKCNE4α—MHC-Clone26，经过酶切鉴定、序列分析证实无误后用BamHI酶切获得转基因表达元件hKCNE4-α—MHC-hGHpA。将hKCNE4-α—MHC-hGHpA显微注入小鼠受精卵雄性原核，并将有活力的受精卵移植到代孕小鼠的输卵管。PCR鉴定出代孕鼠所生小鼠中转基因阳性原代（G0）小鼠，运用RT—PCR-测序技术对阳性G0小鼠与野生型小鼠交配所生的一代（G1）小鼠进行检测以识别出表达转基因的阳性小鼠。结果成功克隆了hKCNE4基因，正确构建了hKCNE4基因心肌特异性表达载体hKCNE4-α—MHC-Clone26。代孕鼠共生产86只仔鼠，9只转基因阳性，2只（分别称其为LineA和LineB）转基因在心脏中高表达。对LineA和LineB的阳性后代进行初步表型分析发现其生长发育良好，繁殖力正常，体表心电图无明显异常。结论成功建立心肌特异性表达hKCNFA转基因小鼠模型，为进一步研究hKCNFA基因对特定类型心房颤动的防治作用提供了一种工具。

英文摘要：

Objective The investigation aimed at producing transgenic mouse model with myocardium-specific overexpression of human KCNFA （hKCNE4） gene so as to research into the prophylactic and therapeutic effect of hKCNE4 gene on a certain subset of atrial fibrillation. Methods hKCNE 4 gene was cloned by reverse transcription-polymerase chain reaction （RT-PCR）, digested with restrictive endonuclease, and inserted into the cardiomyocyte-specific expressing plasmida-MHC-Clone 26 by ligase. Enzymatic and sequencing analyses were performed to confirm hKCNE4-α-MHC- Clone 26 prior to obtaining the BamHI treated transgenic element hKCNE4-α-MHC-hGH pA. The hKCNE4-α-MHC- hGH pA was micminjected into the male pronucleus of fertilized oocyte of C,57BL mice and the oocytes surviving micminjection were transplanted into the oviducts of pseudo-pregnant mice. The positive transgenic founder （Go generation） mice were identified by PCR from offspring generated by pseudo-pregnant mice. The Go transgenic mice were mated respectively with wild-type C57BL mice to reproduce G1 transgenic progeny. RT-PCR and sequencing were applied to screening for transgenic mice expressing hKCNE4 from G1 transgenic descendants. Results hKCNE4 gene was cloned and ligated to the vectora-MHC-Clone 26 correctly. 86 mice were generated 9 of which were identified to be transgenic and the transgene hKCNE 4 carried by 2 transgenic mice （titled as Line A and Line B respectively） was not only transmitted to the next generation but also overexpressed restrictedly in the hearts. Preliminary phenotypic analysis on the transgenic progeny of Line A and Line B showed that they developed normally, reproduced generally, and suffered from no arrhythmia documented by surface electrocardiography. Conclusion The results indicate that the transgenic mice cardiomyocytespecifically overexpressing hKCNE 4 are successfully procured, which provides a valuable tool for the exploration of the preventive and remedial influence of hKCNE 4 on a subgroup of atrial fibrillati