小菜蛾颗粒体病毒DNA限制性酶切分析及基因组DNA片段克隆

小菜蛾颗粒体病毒DNA限制性酶切分析及基因组DNA片段克隆 THE ANALYSIS OF RESTRICTION ENZYME OF PLUTELLA XYLOSTELLA GRANULOSIS VIRUS DNA AND CLONING OF DNA FRAGMENTS 孟小林 Meng Xiaolin 1 first-author 肖红 Xiao Hong 1 吕颂雅 Lu Songya 1 徐进平 Xu Jinpin 1 叶林柏 Ye Linbai 1 ＳｕｚａｎｎｅＭ．Ｔｈｉｅｍ 1 武汉大学病毒研究所 武汉大学病毒研究所 Institute of Virology, Wuhan University, Wuhan 430072,China) Suzanne M.Thiem Institute of Virology, Wuhan University, Wuhan 430072,China) Suzanne M.Thiem 小菜蛾颗粒体病毒ＤＮＡ用ＢａｍＨＩ和ＸｈｏＩ酶切，经０．７５％琼脂糖凝胶电泳，分别得到１２条带和８条带，平均分子量为１１３．０ｋｂ．用Ｓｕｐｅｒｃｏｓ１Ｃｏｓｍｉｄ作载体，将Ｓａｕ３ＡＩ部分消化的ＰｘＧＶ－ＤＮＡ随机片段插入该载体的ＢａｍＨＩ酶切位点，转化于ＸＬ１－Ｂｌｕｅ受体菌，经Ａｍｐ平板筛选转化子，挑出２５６个Ａｍｐ＋菌落，以３２Ｐ－ｄＣＴＰ标记的ＰｘＧＶ－ＤＮＡ为探针，斑点杂交筛选得到５９个阳性重组体，再经电泳检测，得到了５９个不同大小的ＰｘＧＶ－ＤＮＡ片段克隆． Plutella xylostella Granulosis Vinus（PxGV）DNA was cut by BamHI and Xhol and electrophoresed in 0.75% agarose gel, producing 12 and 8 fragments respectively. The average molecular weight of PxGV DNA 113.0 kb. PxGV DNA was digested partially by Sau3Al, fragments were recombinants with Supercos 1 cosmid vector and transformated in the XL1 Blue.256 transformants were selected from ampicilin plate,59 positive recombinans were selected by dot blot using genome PxGV DNA 32 P dCTP labelled as probe and constructed 59 different size PxGV DNA fragments colone by the examination of agarose gel. 小菜蛾颗粒体病毒 酶切图谱 DNA片段克隆 Plutella xylostella granulosis virus restrction map of PxGV DNA DNA fragment clone Q78 国家自然科学基金(61672102) 1997-06-01 2021-04-01 6