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

人干细胞因子cDNA的设计、合成与克隆 被引量：6

The design,chemical synthesis and cloning of the human stem cell factor cDNA
下载PDF
导出
摘要 选择大肠杆菌的偏爱密码子,分成18个长约60个碱基且相互配对的寡核苷酸片段,合成了可溶形式的人干细胞因子(hSCF)的cDNA。合成片段经纯化、5'-端磷酸化后,通过PCR进行合成hSCF的cDNA一次性组装与扩增,得到了含有可溶形式的hSCF的全部编码区及EcoRⅠ、SalⅠ位点在内的全长共515个碱基对的DNA片段。回收该DNA片段,酶切消化,定向克隆进质粒pUC19。经酶切鉴定以及序列测定,得到了全序列正确的克隆株,为下一步的工作奠定了基础。 By selectingthehigh-usagecodonsin Escherichia coli,thesolubleformhSCFcDNA,consistof515base-pairs(bp),was dividedinto18oligonucleotidefragmentsandsynthesizedon ABI381AtypeDNAsynthesizer.Thesynthetic18fragmentswereassembledin onlyonestepby a PCRapproachandinsertedintopUC19for cloning.Thesequenceof the wholesyntheticgenewasconfirmedby DNAsequencing.
作者 赵志虎 张京生 曹诚 马清钧
机构地区 军事医学科学院生物工程研究所
出处 《生物技术通讯》 CAS 2002年第1期26-29,共4页 Letters in Biotechnology
关键词 人干细胞因子 基因合成 聚合酶链式反应 CDNA 设计 克隆 hSCF genechemicalsynthesis PCR
分类号 Q781 [生物学—分子生物学] R394.8 [医药卫生—医学遗传学]
  • 相关文献

参考文献12

  • 1Williams DE, Eisenman J, Baird A, et al. Identification ora ligand for the c-kit proto-oncogene[J]. Cell, 1990, 63(1):167
  • 2Copeland NG, Gilbert DJ, Cho BC, et al. Mast cell growth factor maps near the steel locus on mouse chromosome 10 and is deleted in a number of steel alleles[J]. Cell, 1990, 63(1):175
  • 3Martin FH, Suggs SV, Langley KE, et al. Primary structure and functional expression of rat and human stem cell factor DNAs [J]. Cell,1990, 63(1 ):203
  • 4Matsui Y, Zsebo KM, Hogan BL. Embryonic expression of a haematopoietic growth factor encoded by the SI locus and the ligand for c-kit[J]. Nature, 1990, 347(6294):667
  • 5Zsebo KM, Wypych J, McNiece IK, et al. Identification, purification, and biological characterization of hematopoietic stem cell factor from buffalo rat liver-conditioned medium[J]. Cell, 1990, 63(1): 195
  • 6Zsebo KM, Williams DA, Geissler EN, et al. Stem cell factor is encoded at the S1 locus of the mouse and is the ligand for the c-kit tyrosine kinase receptor[J]. Cell, 1990, 63(1):213
  • 7Huang E, Nocka K, Beier DR, et al. The hematopoietic growth factor KL is encoded by the SI locus and is the ligand of the c-kit receptor,the gene product of the W locus[J]. Cell, 1990, 63(1):225
  • 8Anderson DM, Lyman SD, Baird A, et al. Molecular cloning of mast cell growth factor, a hematopoietin that is active in both membrane bound and soluble forms[J]. Cell, 1990, 63(1 ):235
  • 9Langley KE, Wypych J, Mendiaz EA, et al. Purification and characterization of soluble forms of human and rat stem cell factor recombinantly expressed by Escherichia coli and by Chinese hamster ovary cells [J]. Arch Biochem Biophys, 1992, 295(1 ):21
  • 10Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning A laboratory manual, 2nd ed, CSHL Press, New York. 1989

同被引文献73

  • 1郭斯启,奚永志,崔建武,习彩霞,刘楠,梁飞,孔繁华.重组人干细胞因子在原核系统中高效表达的策略[J].中国输血杂志,2004,17(4):227-231. 被引量：3
  • 2J萨姆布鲁克.分子克隆实验指南[M].北京:科学出版社,1992..
  • 3姚泉洪 黄晓敏 彭日荷等.连续延伸法合成解氨酶基因及测序[J].上海农业学报,1999,15:11-11.
  • 4http://tools.neb.com/inbase/list.php.
  • 5Wu H,Hu Z,Liu X Q.Protein trans-splicing by a split intein encoded in a split DnaE gene of Synechocystis sp.PCC6803[J].Proc Natl Acad Sci USA,1998,95(16):9226-9231.
  • 6Muralidharan V,Muir T M.Protein ligation:an enabling technology for the biophysical analysis of proteins[J].Nat Meth,2006,3(6):429-438.
  • 7Volkmann G,Murphy P W,Rowland E E,et al.Intein-mediated cyclization of bacterial acyl carrier protein stabilizes its folded conformation but does not abolish function[J].J Biol Chem,2010,285(12):8605-8614.
  • 8Tavassoli A,Benkovic S J,Split-intein mediated circular ligation used in the synthesis of cyclic peptide libraries in E.coli[J].Nat Protoc,2007,2(5):1126-1133.
  • 9Kritzer J A,Hamamichi S,McCaffery J M,et al.Rapid selection of cyclic peptides that reduce α-synuclein toxicity in yeast and animal models[J].Nat Chem Biol,2009,9(5):655-663.
  • 10Naumann T A,Tavassoli A,Benkovic S J.Genetic selection of cyclic peptide Dam methyltransferase inhibitors[J].Chembiochem,2008,9(2):194-197.

引证文献6

二级引证文献10

生物技术通讯
2002年 第1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