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人野生型抑癌基因PTEN/MMAC1的巢式PCR法克隆、测序及表达载体的构建 被引量：2

Cloning and Sequencing of the Human Tumor Suppressor Gene PTEN/MMAC1 by RT-nested PCR and Construction of its Expression Vector
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摘要 目的 :克隆人野生型抑癌基因 PTEN/MMAC1 c DNA序列并构建其表达载体。方法 :利用RT- nested PCR法从正常人胎盘组织中扩增出约 1 2 0 0 bp的 DNA片段 ,与 p UCm- T载体连接 ,作全自动测序确证 ,并将其重组入 pc DNA 3.1载体中 ,构建为表达质粒 pc DNA- w P。结果 :对 PCR产物进行测序 ,序列基本正确。结论 :利用 RT- nested PCR法成功克隆了人野生型抑癌基因 PTEN/MMAC1c DNA序列并构建了表达质粒 pc DNA- w P。 Objective: To clone the cDNA of the human tumor suppressor gene PTEN/MMAC1 and construct its expression vector. Methods: About 1 200 bp DNA fragment was amplified from human placenta tissues by using RT nested PCR and was cloned into pUCm T vector after automatic sequenced, then the fragment was linked with expression vector pcDNA 3.1. Results:The PTEN/MMAC1 cDNA had been cloned correctly and its expression vector pcDNA wP also had been constructed. Conclusion: Human tumor suppressor gene-PTEN/MMAC1 cDNA had been cloned by RT nested PCR and its expression vector had been constructed successfully. The result makes the base for further study on the tumor suppression function of PTEN/MMAC1.
作者 田梅 刘林林 李修义
机构地区 吉林大学公共卫生学院卫生部放射生物学重点实验室 吉林大学第二医院放疗科
出处 《吉林大学学报（医学版）》 CAS CSCD 北大核心 2003年第1期9-12,共4页 Journal of Jilin University：Medicine Edition
基金 国家自然科学基金资助课题 (39970 2 2 9)
关键词 PTEN/MMAC1 逆转录聚合酶链反应 巢式PCR法 表达载体 基因 抑制 肿瘤 基因表达 PTEN/MMAC1 reverse transcriptase polymerase chain reaction RT nested PCR expression vector genes,suppressor,tumor gene expression
分类号 Q782 [生物学—分子生物学]
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参考文献6

  • 1[1]Steck PA, Pershouse MA, Jasser SA, et al. Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers[J]. Nat Genet,1997,15:356-362.
  • 2[2]Myers MP, Stolarov JP, Eng C, et al. P-TEN, the tumor suppressor from human chromosome 10q23, is a dual-specificity phosphatase[J]. Proc Nat l Acad Sci USA, 1997,94:9052-9057.
  • 3[3]Whang YE, Wu X, Suzuki H, et al. Inactivation of the tumor suppr essor PTEN/MMAC1 in advanced human prostate cancer through loss of expression[J ]. Proc Natl Acad Sci USA, 1998,95:5246-5250.
  • 4[4]Cantley LC, Neel BG. New insights into tumor suppression: PTEN suppresses tumor formation by restraining the phosphoinositide 3-kinase/AKT path way[J]. Proc Natl Acad Sci USA, 1999,96:4240-4245.
  • 5[5]Persad S, Attwell S, Gray V, et al. Inhibition of integrin-linked kinase (ILK) suppresses activation of protein kinase B/Akt and induces cell cycle arrest and apoptosis of PTEN-mutant prostate cancer cells[J]. Proc Natl Acad Sci USA, 2000,97:3207-3212.
  • 6[6]Wu X, Senechal K, Neshat MS, et al. The PTEN/MMAC1 tumor suppressor phosphatase functions as a negative regulator of the phosphoinositide 3-kin ase/Akt pathway[J]. Proc Natl Acad Sci USA, 1998,95:15587-15591.

同被引文献22

  • 1Roth JA, Cristianoo RJ. Gene therapy for cancer:what have we done and where are we going? [J]. J Natal Cancer Inst, 1997,89: 21-39.
  • 2Weichselbaum RR, Hallanhan DE, Becktt MA, et al. Gene therapy by radiation prefesentially radiosensitizes tumor cell[J]. Cancer Res, 1994, 54: 4266-4269.
  • 3Scott SD, Marples B, Hendry JH, et al. A radiation-controlled molecular switch for use in gene therapy of cancer [J]. Gene Ther, 2000, 7 (13): 1121-1125.
  • 4Steck PA, Pershouse MA, Jasser SA, et al. Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23. 3 that is mutated in multiple advanced cancers [J]. Nat Genet, 1997, 15: 356-362.
  • 5Persad S, Attwell S, Gray V, et al. Inhibition of intergrinlinked kinase (ILK) suppresses activation of protein kinase B/AKT and induces cell cycle arrest and apoptosis of PTEN-mutant prostate cancer cells [J]. Proc Natl Acad Sci U S A,2000, 97 (7): 3207-3212.
  • 6Weng LP, Brown JL, Eng C. PTEN induces apoptosis and cell cycle arrest theough phoaphoinositol-3-kinase/AKT-dependent and independent pathways [J]. Hum Mol Genet, 2001,10:237-242.
  • 7Huang PH, Hokeun, Kim DS. Supperssion of tumorigenicity and metastasis in B16F10 cells by PTEN/MMAC1/TEP1 gene [J]. Cancer Lett, 2001, 172 (1):83.
  • 8Stewart AL, Mhashilkar AM, Yang XH, et al. PI3Kinase blockade by ad-PTEN inhibits invision and induces apoptosis in RGP and metastasis melanoma cells [J]. Mol Med, 2002,8 (8):451.
  • 9Li J,Yen C,Liaw D,et al.PTEN,a putative protein tyrosine phosphatase gene mutated in human brain, breast and prostate cancer [J].Science,1997,275(5308):1 943～1 947.
  • 10Steck P A, Pershouse M A, Jasser S A, et al. Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers [J].Nat Genet,1997,15(4):356～362.

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吉林大学学报（医学版）
2003年 第1期

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