期刊文献+

Preparation of high molecular weight (HMW) genomic DNA from tea plant (Camellia sinensis) for BAC library construction 被引量:1

Preparation of high molecular weight (HMW) genomic DNA from tea plant (Camellia sinensis) for BAC library construction
下载PDF
导出
摘要 A bacterial artificial chromosome (BAC) library is an invaluable resource tool to initiate tea plant genomics research, and the preparation of high molecular weight (HMW) genomic DNA is a crucial first step for constructing a BAC Library. In order to construct a BAC library for enhancing tea plant genomics research, a new method for the preparation of tea pant high molecular weight (HMW) genomic DNA must be developed due to young tea plant leaves and shoots are notably rich in both tea polyphenols and tea polysaccharides. In this paper, a modified method for preparing high quality tea plant HMW genomi~ DNA was optimized, and the quality of tea plant genomic DNA was evaluated. The results were as follows: Critical indicators of HMW DNA preparation were the appearance of the smooth nuclei in solution (as opposed to sticky-gummy) before agarose plug solidification, non-dark colored nuclei plugs after lysis with an SDS/proteinase K solution, and the quality and quantity of HMW DNA fragments after restriction enzyme digestion. Importantly, 1% dissolved PVP-40 and 1% un-dissolved PVP-40 during the nuclei extraction steps, in conjunction with the removal of PVP-40 from the plug washing and nuclei lysis steps, were critical for achieving HWM tea plant DNA suitable for BAC library construction. Additionally, a third PFGE fraction selection step to eliminate contaminating small DNA fragments. The modifications provided parameters that may have prevented deleterious interactions from tea polyphenols and tea polysaccharides. The HMW genomic DNA produced by this new modified method has been used to successfully construct a large-insert tea plant BAC library, and thus may be suitable for BAC library construction from other plant species that contain similarly interfering compounds.
出处 《Journal of Agricultural Science and Technology》 2009年第1期1-10,共10页 农业科学与技术(美国大卫英文)
关键词 tea plant bacterial artificial chromosome library BAC clone tea polyphenols high molecular weight genomic DNA preparation Camellia sinensis 基因组DNA BAC文库 超高分子量 幼龄茶树 HMW C库 制备 细菌人工染色体
  • 相关文献

参考文献9

二级参考文献101

  • 1骆耀平,董尚胜,童启庆,夏涛,屠幼英,须海荣.7个茶树品种新梢生育过程中β-葡萄糖苷酶活性变化[J].茶叶科学,1997,17(S1):25-28. 被引量:44
  • 2陈宗懋.茶叶中的农药残留最高允许限量(MRL)[J].农药科学与管理,1992,13(4):1-5. 被引量:5
  • 3夏涛,童启庆,董尚胜,骆耀平.红茶萎凋发酵中β-葡萄糖苷酶的活性变化[J].茶叶科学,1996,16(1):63-66. 被引量:70
  • 4[1]Chen L.,Zhao L.P.,and Gao Q.K.,2005,Generation and analysis of expressed sequence tags from the tender shoot cDNA library of tea plant (Camellia sinensis),Plant Sci.,168:359-363
  • 5[2]Edwards J.W.,Walker E.L.,and Coruzzi G.M.,1990,Cell-specific expression in transgenic plants reveals nonoverlapping roles for chloroplast and cytosolic glutamine synthetase,Proc.Natl.Acad.Sci.,87:3459-3463
  • 6[4]Kato M.,Mizuno K.,Crozier A.,Fujimura T.,and Ashihara A.,2000,Caffeine synthase gene from tea leaves,Nature,406:956-957
  • 7[5]Kato M.,Mizuno K.,Fujimura T.,Iwama M.,Irie M.,Corzier A.,and Ashihara H.,1999,Purification and characterization of caffeine synthase from tea leaves,Plant Physiol.,120:579-586
  • 8[8]Matsumoto S.,Takeuchi A.,and Hayatsu M.,1994,Molecular cloning of phenylalanine ammonia-lyase cDNA and classification of varieties and cultivars of tea plants (Camellia sinensis) using the tea PAL cDNA probe,Theor.Appl.Genet.,89:671-675
  • 9[9]Mizutani M.,Nakanishi H.,Ema J.,Ma S.J.,Noguchi E.,Inohara-Ochiai M.,Fukuchi-Mizutani M.,Nakao M.,and Sakata K.,2002,Cloning of β-primeverosidase from tea leaves,a key enzyme in tea aroma formation,Plant Physiol.,130:2164-2176
  • 10[10]Mondal T.K.,Bhattacharya A.,Ahuja P.S.,and Chand P.,2001,Transgenic tea[Camellia sinensis (L.) O.Kuntze cv.Kangral Jat] plants obtained by Agrobacterium-mediated transformation of somatic embryos,Plant Cell Rep.,20 (8):712-720

共引文献168

同被引文献17

引证文献1

二级引证文献6

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