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构建携带组织金属蛋白酶抑制物ＴＩＭＰ－１基因的逆转录病毒载体，并用于感染胰腺癌ＳＷ１９９０细胞系，观察其侵袭、转移能力的变化．利用常规分子克隆技术，通过对逆转录病毒表达载体ＰＭＮＳＭ（Ｆ６）质粒的去磷酸化、粘端连接，构建ＴＩＭＰ－１基因逆转录病毒表达载体ＰＭＮＳＭ／ｈＴＩＭＰ；经限制性内切酶酶切、电泳鉴定其结构正确后，用电穿孔法对体外培养的ＰＡ１３７包装细胞进行基因转移，Ｇ４１８筛选阳性克隆，收集上清，并用于感染胰腺癌ＳＷ１９９０细胞系的研究．构建获得ＴＩＭＰ－１基因逆转录病毒表达载体ＰＭＮＳＭ／ｈＴＩＭＰ，有效滴度为１０４ＣＦＵ／ｍＬ～１０６ＣＦＵ／ｍＬ．本研究构建获得的ＴＩＭＰ－１基因逆转录病毒表达载体，在感染胰腺癌ＳＷ１９９０细胞系，观察其侵袭、转移能力的研究中得到有效应用
A retroviral vector carrying the metalloproteinase inhibitor TIMP-1 gene was constructed and used to infect pancreatic cancer SW1990 cell line to observe the changes of its invasion and metastasis capacity. Using conventional molecular cloning techniques, the retroviral expression vector PMNSM/hTIMP of the retroviral expression vector PMNSM (F6) was constructed by dephosphorylation and sticky-end ligation of the retroviral expression vector PMNSM (F6); After electrophoresis was used to identify the correct structure, the PA137 packaging cells were cultured in vitro by electroporation. The positive clones were selected by G418, and the supernatant was collected and used to infect pancreatic cancer SW1990 cell line. The retroviral expression vector PMNSM/hTIMP of TIMP-1 gene was constructed and the effective titer was 104 CFU/mL to 106 CFU/mL. In this study, the retroviral expression vector of TIMP-1 gene was constructed and used in the study of pancreatic cancer SW1990 cell line to observe its invasion and metastasis ability.