單純皰疹病毒Ⅱ型胸苷激酶基因及血管生成抑制素融合基因真核表達質粒的構建及鑒定_《中國普外基礎與臨床雜志》

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昆明醫學院第二附屬醫院肝膽外科（昆明650101）;

關鍵詞：

單純皰疹病毒Ⅱ型胸苷激酶基因血管抑制因子克隆真核表達載體融合基因

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目的對單純皰疹病毒Ⅱ型胸苷激酶基因及血管生成抑制素進行基因擴增，克隆構建真核表達質粒pcDNA3/HSVⅡ TK/As。方法從單純皰疹病毒Ⅱ型（HSVⅡ）Sav株感染的Hep2細胞上清液中提取HSV2基因組DNA，以此為模板，用PCR方法擴增胸苷激酶（TK）基因，將擴增的片段克隆入載體pcDNA3中，挑取陽性克隆測序。限制性核酸內切酶酶切pcDNA3/HSVⅡTK,得到目的基因TK，將其克隆于已構建的真核表達載體pcDNA3/As上。結果HSVⅡTK基因全部編碼區為1 128 bp，基因序列與genebank中報道相符。新質粒pcDNA3/HSVⅡTK/As經限制性核酸內切酶(BamH Ⅰ,Hind Ⅲ)酶切后得到700 bp(As)及1000 bp(HSVⅡTK)相應基因片段。結論本研究成功構建pcDNA3/HSVⅡTK/As真核表達質粒。

引用本文： 王琳,郭永章,李立,張小文,李曉,崔江云. 單純皰疹病毒Ⅱ型胸苷激酶基因及血管生成抑制素融合基因真核表達質粒的構建及鑒定. 中國普外基礎與臨床雜志, 2002, 9(4): 246-248. doi: 復制

1.	Moolten FL.Tumor chemosensitivity conferred by inserted herpes thymidine kinase genes: paradigm for a prospective cancer control strategy [J]. Cancer Res,1986； 46(10)∶5276.
2.	Nagy H, Panis Y, Fabre M, et al.Are hepatomas a good target for suicide gene therapy? An experimental study in rats using retroviralmediated transfer of thymidine kinase gene [J]. Surgery,1998; 123(1)∶ 19.
3.	O’Reilly MS, Holmgren L, Shing Y,et al.Angiostatin: a novel angiogenesis inhibitor that mediates the suppression of metastasis by a Lewis lung carcinoma [J]. Cell,1994; 79(2)∶ 315.
4.	Kit S, Kit M, Qavi H, et al. Nucleotide sequence of the herpes simplex virus type 2(HSV2)thymidine kinase gene and predicted amino acid sequence of thymidine kinase polypeptide and its comparison with the HSV1 thymidine kinase gene [J]. Biochim Biophys Acta,1983; 741(2)∶158.
5.	薩姆布魯克J，弗里奇EF，曼尼阿蒂斯文T.分子克隆實驗技術指南[M]. 第2版.北京：科學出版社，1992∶237~340.
6.	Kuriyama S, Nakatani T, Masui K, et al. Bystander effect caused by suicide gene expression indicates the feasibility of gene therapy for hepatocellular carcinoma [J]. Hepatology,1995; 22(6)∶1838.
7.	Gately S,Twardowski P,Stack MS,et al.The mechanism of cancermediated conversion of plasminogen to the angiogenesis inhibitor angiostatin [J]. Proc Natl Acad Sci USA,1997; 94(20)∶10868.
8.	Sim BK,O’Reilly MS,Liang H,et al. A recombinant human angiostatin protein inhibits experimental primary and metastatic cancer [J]. Cancer Res, 1997; 57(7)∶1329.

1. Moolten FL.Tumor chemosensitivity conferred by inserted herpes thymidine kinase genes: paradigm for a prospective cancer control strategy [J]. Cancer Res,1986； 46(10)∶5276.
2. Nagy H, Panis Y, Fabre M, et al.Are hepatomas a good target for suicide gene therapy? An experimental study in rats using retroviralmediated transfer of thymidine kinase gene [J]. Surgery,1998; 123(1)∶ 19.
3. O’Reilly MS, Holmgren L, Shing Y,et al.Angiostatin: a novel angiogenesis inhibitor that mediates the suppression of metastasis by a Lewis lung carcinoma [J]. Cell,1994; 79(2)∶ 315.
4. Kit S, Kit M, Qavi H, et al. Nucleotide sequence of the herpes simplex virus type 2(HSV2)thymidine kinase gene and predicted amino acid sequence of thymidine kinase polypeptide and its comparison with the HSV1 thymidine kinase gene [J]. Biochim Biophys Acta,1983; 741(2)∶158.
5. 薩姆布魯克J，弗里奇EF，曼尼阿蒂斯文T.分子克隆實驗技術指南[M]. 第2版.北京：科學出版社，1992∶237~340.
6. Kuriyama S, Nakatani T, Masui K, et al. Bystander effect caused by suicide gene expression indicates the feasibility of gene therapy for hepatocellular carcinoma [J]. Hepatology,1995; 22(6)∶1838.
7. Gately S,Twardowski P,Stack MS,et al.The mechanism of cancermediated conversion of plasminogen to the angiogenesis inhibitor angiostatin [J]. Proc Natl Acad Sci USA,1997; 94(20)∶10868.
8. Sim BK,O’Reilly MS,Liang H,et al. A recombinant human angiostatin protein inhibits experimental primary and metastatic cancer [J]. Cancer Res, 1997; 57(7)∶1329.

《中國普外基礎與臨床雜志》

單純皰疹病毒Ⅱ型胸苷激酶基因及血管生成抑制素融合基因真核表達質粒的構建及鑒定

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《中國普外基礎與臨床雜志》

單純皰疹病毒Ⅱ型胸苷激酶基因及血管生成抑制素融合基因真核表達質粒的構建及鑒定

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

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