草菇ras启动子区域的克隆及其序列分析
Cloning and Sequence Analysis of ras Promoter Region from Volvariella volvacea
刘世强 杨丽卿 郭丽琼 林俊芳
摘 要:根据Kajiwara S[1]等报道的ras启动子的序列设计并合成一对特异引物,以草菇菌丝的总DNA为模板,通过PCR扩增,获得一条长约0.75kb片段.DNA序列测定结果表明,该片段长度为751bp.采用Blast软件和Promoter predictions软件进行启动子序列结构分析,结果表明,该片段中243~293 bp和539~589 bp两区域为ras启动子的基础启动子区.在283bp和579bp处有两个转录起始位点,在244~247bp和292~295bp之间有2个TATA box,在36~39bp、377~380bp、434~437bp和716~719bp之间有4个CAAT box.此外,该片段中还含有9个G box、12个Ⅰ box、2个P box以及3个重要顺式作用元件:ABRE元件、WUN-motif(基元)和HSE元件.
关键词:草菇; ras启动子; PCR扩增; 序列分析
分类号:S646.130.6 文献标识码:A
文章编号:1005-9873(2004)01-0001-06
基金项目:国家自然科学基金资助项目(30060054和30371000)、广东省自然科学基金资助项目(032239)和华南农业大学校长基金项目(5100-k03003)的部分研究内容
作者简介:刘世强(1978-),男,华南热带农业大学农学院在读硕士研究生,主要从事食用菌的基因工程研究与开发.林俊芳,本文通讯作者
作者单位:刘世强(中国热带农业科学院热带作物生物技术国家重点实验室,海口,571701)
杨丽卿(中国热带农业科学院热带作物生物技术国家重点实验室,海口,571701)
郭丽琼(中国热带农业科学院热带作物生物技术国家重点实验室,海口,571701;华南农业大学食品学院生物工程系,广州,510640)
林俊芳(中国热带农业科学院热带作物生物技术国家重点实验室,海口,571701;华南农业大学食品学院生物工程系,广州,510640)
参考文献:
[1]Kajiwara S,Shishido K.Characterization of the promoter region of the basidiomycete Lentinus edodes Le.ras gene[J].FEMS Microbiology letters,1992,92(2):147~150.
[2]Katsutoshi Hori, Susumu Kajiwara, Takeshi Saito, et al. Cloning, sequence analysis and transcriptional expression of a ras gene of the edible basidiomycete Lentinus edodes[J]. Gene, 1991,(105):91~96.
[3]Hall A , Brown R. Human N-ras: cDNA cloning and gene structure[J]. Nucleic Acids Res,1985, 13(14):5255~5268.
[4]Myers RM,Tilly K,Maniatis T.Fine structure genetic analysis of a beta-globin promoter[J].Science,1986, 232(4750):613~618.
[5]Kadonaga JT,Carner KR, Masiarz FR, et al.Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain[J].Cell,1987, 51(6):1079~1090.
[6]Bishop JG, Corces VG. Expression of an activated ras gene causes developmental abnormalities in transgenic Drosophila melanogaster[J]. Gene Dev,1988, 2(5):567~577.
[7]郭丽琼,陈守才,林俊芳.食用菌遗传转化研究进展[J].食用菌学报,2001,8(4):47~53.
[8]Yanai K, Yonekura K, Usami H,et al. The integrative transformation of Pleurotus ostreatus using Bialaphos resistance as a dominant selectable marker[J]. Bioscience Biotechnology and Biochemistry,1996,60(3):472~475.
[9]Granado JD,Kertes-Chaloupkova, Aebi M, et al. Restriction enzyme-mediated DNA integration in Coprinus cinereus[J].Molecular and General Genetics,1997,256(1):28~36.
[10]Sato T,Yaegashi K, Ishll S, et al. Transformation of edible basidiomycete Lentinus edodes by restriction enzyme-mediated DNA integration of plasmid DNA[J]. Biosci. Biotechnol. Biochem.,1998,62(12):2346~2358.
[11]Ogawa K, Yamazaki T, Hasebe T,et al. Molecular breeding of the basidiomycete Coprinus cinereus strains with high lignin-decolorization and degradation activities using novel heterologous protein expression vectors[J]. Applied Microbiology and Biotechnology,1998,49(3):285~289.
[12]Joseph Sambrook, David W Russel(黄培堂译).分子克隆实验指南[M].北京:科学出版社,2002.26~27 ,96~99.
[13]孙晓红,陈明杰,潘迎捷.启动子克隆概述[J].食用菌学报,2002,9(3):57~62.
[14]Chan SJ,Oliva AA, La Mendola J, et al. Conservation of the prohormone convertase gene family in metazoa:analysis of cDNA encoding a PC3-like protein from hydra[J]. Proc Natl Acad Sci USA,1992,89(15):6678~6682.
[15]张晓宁,王昊,陈火英,等.蓝藻启动子活性片段的克隆及其序列分析[J].复旦大学学报(自然科学版),2002,41(2):227~229.
[16]陈明杰,汪昭月,贺冬梅,等.低温影响草菇蛋白质组分变化的研究[J].食用菌学报,1995,2(4):28~31.
[17]张树庭.草菇[M].香港:香港中文大学出版社,1978.143~147.
[18]Chang ST, Hayes WA.Volvariella volvacea in the biology and cultivation of edible fungi[M]. New York:Academic Press ,1978.573~605.
[19]Chang ST, Quimil TH. Tropical mushrooms biological nature and cultivation methods. Hong Kong: The Chinese University Press,1982.69~80.
[20]王富民,宫秀荣.草菇保鲜生理研究[J].食用菌,1990,12(6):37~38.
[21]王富民,宫秀荣.草菇保鲜及有关生理变化[J].上海农业学报,1992,8(3):60~66.
[22]段学武,庞学群,张昭其.草菇低温贮藏及有关生理变化研究[J].热带作物学报,2000, 21(4):75~79.
[23]郑国扬, 廖汉泉.中国草菇生产[M].北京:中国农业出版社,2000.12~25.