pXZ208-EGFP—HO-1慢病毒表达载体的构建与鉴定

Construction and identification of recombinant lentivirus vector carrying HO-1

目的构建大鼠血红素加氧酶-1（HO-1）的慢病毒表达载体。方法应用基因重组技术和限制性内切酶酶切构建并鉴定pXZ208-EGFP—HO-1表达载体，脂质体共转染法在包装细胞293FT中包装含有HO-1的慢病毒再感染大鼠原代心肌细胞。结果酶切出两条分别为egfp—ho-1片段大小约1640bp、pXZ208片段大小约为8583bp，酶切鉴定正确，包装出的慢病毒成功感染心肌细胞，感染效率可达60％以上。结论成功构建pXZ208-EGFP—HO-1的慢病毒表达载体，并建立高效稳定表达egfp—ho-1的慢病毒转染系统。

Objective To construct a lentiviral expression vector encoding rat heme oxygenase -1 （ HO-1 ） and EGFP genes. Methods A lentiviral expression vector for EGFP-HO-1 was constructed by using recombinant DNA technique and identified by digestion with restriction enzyme. By liposome-mediated transfection, lentiviral particles expressing EGFP-HO-1 in packaging cells （293FF cells） were generated and transfected into 293FT cells. Results The length of EGFP-HO-1 gene and pXZ208 was 1640 bp and 8583 bp respectively, which was confirmed by enzyme digestion. There lentiviral particles were infected into myocytes successfully with the efficiency being above 60%. Conclusion The lentiviral expression vector pXZ208-EGFP-HO-1 was successfully generated, and an efficient and stable lentiviral system expressing EGFP-HO-1 was constructed.