期刊文献+

西农萨能奶山羊β酪蛋白基因5′侧序列通用表达载体的构建 被引量:2

Universal expression vector construction of beta-casein gene 5′ flanking sequence from Xinong Saanen Dairy Goat
下载PDF
导出
摘要 目的:检测β酪蛋白基因5′侧序列的启动BglⅡ效果及构建真核表达载体pcDNA3.165.方法:采用SalⅠ和BamHⅠ对pMDT/65双酶切,得到6465bp的β酪蛋白基因5′侧序列;将启动子片段插入到XhoⅠ和BglⅡ双酶切后的pGL3Basic载体里;获得重组报告基因载体pGL3B65.瞬时转染西农萨能奶山羊原代培养的乳腺上皮细胞,检测β酪蛋白基因5′侧序列的启动效果.将β酪蛋白基因5′侧序列插入真核表达载体pcDNA3.1(-)的XhoⅠ和BamHⅠ位点之间,从而获得载体pcDNA3.165.结果:长度为6465bp的β酪蛋白基因5′侧序列能有效地启动荧光素酶报告基因,并正确构建了真核表达载体pcDNA3.165.结论:克隆的β酪蛋白基因5′侧序列在转录水平上具有生物学功能,为进一步建立转基因动物乳腺生物反应器奠定了坚实的基础. AIM: To detect the promote effect of β-casein gene 5′ flanking sequence and to construct universal expression vector pcDNA3.1-65. METHODS: ① After the β-casein gene 5′ flanking sequence with 6465 bp from pMD-T/65 vector was double digested with [WTFX]SalⅠ/ [WTFX]BamHⅠ, the 6465 bp sequence was obtained.And 6465 bp sequence was inserted into pGL3-Basic vector which had been double digested with XhoⅠ/BglⅡ.The new clone was named pGL3-B65.② Detect the activity of pGL3-B65 constructed by transient transfection in primary goat mammary gland epithelial cells.③ Insert the β-casein gene 5′ flanking sequence into [WTFX]XhoⅠ /[WTFX]BamHⅠ of the pcDNA3.1 vector to construct pcDNA3.1-65 vector. RESULTS: The β-casein gene 5′ flanking sequence with 6465 bp sequence could promote luciferase reporter gene effectively. The pcDNA3.1-65 vector was well constructed. CONCLUSION: It indicates that the cloned β-casein gene 5′ flanking sequence would have its biological function at the transcriptional level, and lay a solid foundation for constructing transgenic animals and its mammary gland bioreactor.
出处 《第四军医大学学报》 北大核心 2005年第12期1066-1069,共4页 Journal of the Fourth Military Medical University
关键词 β-酪蛋白基因5’侧序列 西农萨能奶山羊 通用表达载体 beta-casein gene 5′ flanking sequence Xinong Saanen Dairy Goat universal expression vector
  • 相关文献

参考文献7

  • 1Provot C, Persuy MA , Mercier JC . Complete sequence of the ovine beta-casein-encoding gene and interspecies comparison [ J ]. Gene,1995 :154 ( 2 ) :259 - 263.
  • 2Houdebine LM. Transgenic animal bioreactors [ J ]. Transgenic Res,2000 :9(45 ) :305 - 320.
  • 3Kang YK, Lee CS , Chung AS, et al. Prolactin-inducible enhancer activity of the first intron of the bovine beta-casein gene [J].Mol Cells, 1998;8 ( 3 ) :259 - 265.
  • 4Kolb A . The first intron of the murine beta-casein gene contains a functional promoter [ J ]. Biochem Biophys Res Commun,2003 ;306 (4) : 1099 - 1105.
  • 5Voigtlander T , Ganten D , Bader M. Transcriptional regulation of the rat renin gene by regulatory elements in intron 1 [J].Hypertension, 1999 ;33 ( 1 ) : 303 - 311.
  • 6Wang GL, Moore ML, McMillin JB. A region in the first exon/intron of rat carnitine palmitoyltransferase Ibeta is involved in enhancement of basal transcription [ J ]. Biochem, 2002; 362 ( 3 ) :609 -618.
  • 7Surinya KH, Cox TC,May BK. Identication and characterization of a conserved erythroid-speeific enhancer located in intron 8 of the human 5-aminolevulinate synthase 2 gene [ J ]. Biol Chem,1998 ;273 ( 27 ) : 16798 - 16809.

同被引文献18

  • 1夏平安,刘维全,江禹,孙绍光,杨淑艳,王吉贵.家蝇卵黄蛋白基因启动子区的克隆与活性分析[J].Acta Genetica Sinica,2004,31(7):688-694. 被引量:6
  • 2宋红卫,郑月茂,张明烽,唐爽,张涌.山羊BLG基因5′调控序列克隆及其调控GFP基因细胞的表达[J].西北农林科技大学学报(自然科学版),2005,33(5):12-14. 被引量:3
  • 3武雅楠,李建国,曹玉凤,韩醒,钟荣珍.硫在反刍动物营养中的作用及其需要量的研究[J].中国畜牧兽医,2006,33(5):16-19. 被引量:11
  • 4Dunn S M, Keough R A,et al. Regulation of a hair follicle keratin intermediate filament gene promoter[J]. Journal of Cell Science,1998,111:3487-3496.
  • 5Fratini A,Powell B C,Rogers G E. Sequence,expression and evolutionary conservation of a gene encoding a glyeine/tyrosine-rich keratin-associated protein of hair. J Biol Chem, 1993,268 : 4511 -4518.
  • 6Keough R,Powell B, Rogers G. Targeted expression of SV40 T antigen in the hair follicle of transgenic mice produces an aberrant hair phenotype[J].Journal of Cell Science, 1995,108 : 957 -966.
  • 7Powell B C,Rogers G E. The formation and structure of human hair[J]. Birkhuser Verlag, 1997 : 59 - 148.
  • 8Powell B C,Sleigh M J ,Ward K A,et al. Mammalian keratin gene families:organization for genes coding for the B-high-sulphur proteins sheep wool[J].Nucleic Acids Res, 1983,11:5327-5346.
  • 9Sami D, Hung Y, Nigel P, et al. Improved wool production in transgenie sheep expressing insulin-like growth factor 1[J].Nature Biotechnology,1996,14:185-188.
  • 10Aoki N,ho K,Ito M.Isolation and characterization of mouse high-glycine/tyrosine proteins[J].J Biol Chen,1997,272 (48):30512-30518.

引证文献2

二级引证文献17

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