摘要目的：构建间隙连接蛋白43（CX43）过表达载体，并对其进行鉴定。方法用内切酶EcoRI、XbaI双酶切含目的基因的质粒和慢病毒载体，将目的基因与酶切线性化的慢病毒载体进行连接，构建重组慢病毒表达载体pHBLV-CMVIE-IRES-ZsGreen-CX43；将构建的载体先进行酶切鉴定再对其测序分析，对测序正确的重组载体和包装质粒共同转染293FT细胞，进行病毒包装和生产，收集浓缩病毒液后在293FT细胞中测定病毒滴度，并采用实时定量聚合酶链反应（PCR）对CX43基因表达进行鉴定。结果酶切鉴定和测序分析均证明CX43慢病毒载体构建成功，包装慢病毒，收集浓缩后滴度为1×108/ml，实时定量PCR鉴定CX43基因在293FT细胞的表达明显增加。结论成功构建了稳定过表达CX43的慢病毒载体。

abstractsObjective To construct and identify lentiviral vectors carrying the connexin43 (CX43) gene. Methods Plasmids containing the CX43 gene and lentiviral vectors were digested using EcoRI/XbaI restriction enzymes, and the target gene fragments were cloned into the lentival vectors to result in CX43 recombinant lentiviral vectors (pHBLV-CMVIE-IRES-ZsGreen-CX43). CX43 recombinant lentiviral vectors were identified by restriction enzyme digestion and electrophoresis, and sequencing was carried on only for correct vectors after identification. The successfully-constructed CX43 recombinant lentiviral vectors and packaging plasmids were mixed and contransfected into 293FT cell for packaging and producting virus. Then the virus was collecting, concentrated and titrated in 293FT cells. Finally, the expression of CX43 gene was assessed by real-time polymerase chain reaction(PCR). Results The restriction enzyme digestion and electrophoresis and sequencing results proved that CX43 recombinant lentiviral vectors were constructed correctly. Lentiviral concentrated virus suspension titer was 1× 108/ml. The up-regulation expression of CX43 was detected correctly by real-time PCR in 293FT cells. Conclusions Stable lentiviral vectors expressing CX43 gene is constructed successfully.