Kir2.1/ Kir2.3嵌合体的构建与表达

摘要

目的 构建Kir2.1/ Kir2.3通道嵌合体,为进一步研究Kir2.1和Kir2.3通道的调控机制打下基础.方法 利用重叠延伸PCR方法构建不同的Kir2.1/ Kir2.3嵌合体质粒:N1P3C3,N3P1C1,N3P3C1,N1P1C3,NIP3C1,N3P1C3.将不同的嵌合体质粒分别用NheI线性化后,转录为cRNA表达于非洲爪蟾卵母细胞,用双电极电压钳记录电流.结果 不同的Kir2.1/ Kir2.3嵌合体质粒构建成功,在非洲爪蟾卵母细胞上可以记录到电流表达.结论 成功构建Kir2.1/ Kir2.3嵌合体并完成了嵌合体通道的异源性表达和电压钳记录.%Aim To construct Kir2. 1 / Kir2. 3 chimeras, and to lay a basis for a future research. Methods Kir2. 1/ Kir2. 3 chimeras were constructed by over-lap extension PCR including N1P3C3, N3P1C1, N3P3C1, N1P1C3, NIP3C1, and N3P1C3. The chimeras would be expressed in Xenopus oocytes, and the currents of chimeras were recorded by TEVC ( two elec-trodes voltage clamp ). Results The chimeras between Kir2. 1 and Kir2. 3 were constructed successfully; the currents could be recorded and analyzed by TEVC. Conclusion The Kir2. 1/ Kir2. 3 chimeras can be successfully constructed.