人TIMMDC1基因在杆状病毒系统中的表达

Expression of the human TIMMDC1 gene in the baculovirus system

旨在杆状病毒系统中表达TIMMDC1/c3orf1(Translocase of inner mitochondrial membrane domain containing1)基因,为其应用和功能研究奠定基础。将TIMMDC1基因克隆至重组杆状病毒表达系统的载体中,构建重组穿梭质粒,转染Bm-N细胞,Western blotting验证蛋白的表达。结果表明,重组克隆质粒p Fast Bac1-GFP-TIMMDC1的两两酶切结果证实重组克隆质粒构建正确。PCR结果显示重组穿梭质粒Bacmid p Fast Bac1+TIMMDC1+GFP可扩增出目的条带。Bm-N-r Bac TIMMDC1的表达产物经Western blotting分析,分别可见相对分子质量57 k D的特异蛋白条带,表明融合蛋白TIMMDC1+GFP在家蚕Bm细胞中成功的进行了表达。本研究成功在杆状病毒表达系统中表达了TIMMDC1基因,为其临床应用和功能研究奠定了基础。

To express the human TIMMDC1/c3orfl （Translocase of inner mitochondrial membrane domain containing 1 ） gene in the baculovirus system, the TIMMDC1 gene was cloned into a vector in the baculovirus system. The constructed recombinant shuttle plasmids were transfected to Bm-N cells. The expressed protein was identified using Western blot analysis. Results showed that the recombinant cloning plasmid pFastBacl-GFP- TIMMDCI was constructed correctly. Target gene bands were amplified from the recombinant shuttle plasmids for TIMMDC1 using PCR. The protein TIMMDC1 was identified using SDS-PAGE and Westem blot analyses. In conclusion, the TIMMDC! gene was successfully expressed in the baculovirus system, which would lay a foun- dation for the study of the gene function.