期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

多发性骨髓瘤细胞株ARH-77L1逆转录酶基因的克隆及体外表达研究

In vitro Cloning and Expression of L1 Reverse Transcriptase Gene in Human Mult iple Myeloma Cell Line ARH-77
下载PDF
导出
摘要 通过菌落原位分子杂交,从多发性骨髓瘤细胞株ARH-77cDNA文库获得L1逆转录酶基因5'端序列.随后使用3'RACE技术,获得L1逆转录酶基因3'端序列及poly(A)尾.生物信息学分析表明:该L1逆转录酶DNA序列有一长552bp开放性阅读框,编码184个氨基酸残基的多肽链,其相对分子质量约为21kDa.同时将编码L1逆转录酶保守区的开放性阅读框DNA片段与原核表达载体pQE30连接,得到重组原核表达质粒,利用大肠杆菌表达并获得L1逆转录酶融合蛋白. The full length sequence with poly(A) tail of L1 reverse transcript as e gene in human multiple myeloma cell line ARH 77 cDNA library was obtained by in situ hybridization and 3′RACE technique.Bioinformatic analysis shows that L1 reverse transcriptase gene contains an open reading frame with 552 base pairs a nd encodes a polypeptide with 184 amino acids,its relative molecular weight is 2 1 kDa.The Recombinant prokaryotic expression plasmid pQE30 L1/ORF was construct ed and transformed into E.coli JM109.L1 fusion protein was expressed under the i nduction of IPTG.
作者 江元山 石奕武 罗赛群 汤立军 田菁燕 胡维新
机构地区 中南大学生物科学与技术学院分子生物学研究中心中国
出处 《生命科学研究》 CAS CSCD 2005年第2期118-123,共6页 Life Science Research
基金 国家自然科学基金资助项目(39880021) 国家自然科学基金资助项目(30270570) 国家自然科学基金资助项目(30300406)
关键词 多发性骨髓瘤 ARH-77 L1逆转录酶 基因克隆 体外表达 human multiple myeloma ARH 77 L1 reverse transcriptase gene cloning gene expression
分类号 Q786 [生物学—分子生物学]
  • 相关文献

参考文献19

  • 1LEVIN H L. It's prime time for reverse transcriptase [ J ]. Cell.1997, 88(1 ): 5-8.
  • 2KAZAZIAN H H Jr, WONG C, YOUSSOUFIAN H, et al. Haemophilia A resulting from de novo insertion of LI sequences represents a novel mechanism for mutation in man[J].Nature, 1988, 332(6160): 164-166.
  • 3SUMINAGA R, TAKESHIMA Y, YASUDA K, et al. Non-homologous recombination between Alu and L1 repeats caused a 430 kb d eletion in the dystrophin gene: a novel source of genomic instability[J]. J Hum Genet, 2000, 45 (6):331-336.
  • 4MIKI Y, NISHISHO I, HORII A, et al. Disruption of the APC gene by a retrotransposal insertion of LI sequence in a colon cancer[J]. Cancer Res, 1992, 52(3): 643-645.
  • 5HATTORI M, KUHARA S, TAKENAKA O, et al. LI family of repetitive DNA sequences in primates may be derived from a sequence encoding a reverse transeriptase- related protein [ J ].Nature, 1986, 321 (6070): 625-628.
  • 6LIN C H, HSIEH S Y, SHEEN I S, et al. Genome-wide hypomethylation in hepatoeellular carcinogenesis[J]. Cancer Res,2001,61 (10): 4238-4243.
  • 7SINGER M F, KREK V, MCMILLAN J P, et al. LI: a human transposable element[J]. Gene, 1993, 135(1-2): 183-188.
  • 8CLEMENTS A P, SINGER M F. The human LI reverse transcriptase: effect of deletions outside the common reverse transcriptase domain[J]. Nucleic Acids Res, 1998, 26( 15): 3528-3535.
  • 9HOHJOH H, SINGER M F. Sequence-specific single-strand RNA binding protein encoded by the human LI retrotransposon [J]. EMBO J, 1997, 16(19): 6034-6043.
  • 10FENG Q, MORAN J V, KAZAZIAN H H Jr, et al. Human L1retrotransposon encodes a conserved endonuclease required for retrotransposition[J]. Cell, 1996, 87 (5): 905-916.

二级参考文献13

  • 1Hallek M, Bergsagel PL, Anderson KC. Multiple myeloma: Increasing evidence for a muhistep transformation process. Blood,1998, 91:3-21.
  • 2Burk KH, Drewinko B, Turjillo JM, Ahearn MJ. Establishment of a human plasma cell line in vitro. Cancer Res, 1978, 38:2508-2513.
  • 3Nishimura A, Morita M, Nishimura Y, Sugino Y. A rapid and highly efficient method for preparation of competent Escherichia coli cells. Nucleic Acids Res, 1990, 18:6169-6173.
  • 4Gado K, Domjan G, Hegyesi H, Falus A. Role of interleukin-6 in the pathogenesis of multiple myeloma. Cell Biol Int, 2000, 24:195 - 209.
  • 5Chauhan D, Kharbanda S, Ogata A, Urashima M, Teoh G, Robertson M, Kufe DW et al. Interleukin-6 inhibits Fas-induced apoptosis and stress-activated protein kinase activation in multiple myeloma cells. Blood, 1997, 89:227-234.
  • 6Bergsagel PL, Kuehl WM. Chromosome translocations in multiple myeloma. Oncogene, 2001, 20:5611 -5622.
  • 7Bergsagel PL, Chesi M, Nardini E, Brents LA, Kirby SL, Kuehl WM. Promiscuous translocations into immunoglobulin heavy chains witch regions in multiple myeloma. Proc Nail Acad Sci USA,1996, 93:13931 - 13936.
  • 8Nishida K, Tamura A, Nakazawa N, Ueda Y. Abe T, Matsuda F,Kashima K et al. The Ig heavy chain gene is frequently involved in chromosomal translocations in multiple myeloma and plasma cell leukemia as detected by in situ hybridization. Blood, 1997, 90:526 - 534.
  • 9Barbacid M. ras genes. Annu Rev Biochem, 1987, 56:779 -827.
  • 10Takahashi S, Reddy SV, Chirgwin JM, Devlin R, Haipek C, Anderson J, Roodman GD. Cloning and identification of annexin Ⅱ as an autocrine/paracrine factor that increases osteoclast formation and bone resorption. J Biol Chem, 1994, 269 : 28696 - 28701.

共引文献3

生命科学研究
2005年 第2期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部