中国棉铃虫核型多角体病毒几丁质酶基因的定位与克隆

Localization and Cloning of the Chitinase Gene of Heliothis armigera Single Nuclocapsid Nucleopolyhedrovirus

以α３２ＰｄＡＴＰ标记含ＣｆＭＮＰＶ几丁质酶基因的重组质粒为探针，在６８℃条件下对棉铃虫单粒包埋核型多角体病毒（ＨａＳＮＰＶ）进行Ｓｏｕｔｈｅｒｎ杂交，将ＨａＳＮＰＶ的几丁质酶基因分别定位在ＢａｍＨＩＥ、ＢｇｌⅡＥ、ＥｃｏＲＩＧ、ＨｉｎｄⅢＦ、ＸｂａＩＨ、ＢａｍＨＩ＋ＨｉｎｄＩＩＭ和ＢａｍＨＩ＋ＸｂａＩＨ，并以ｐＴＺ１９Ｒ为载体获得了ＸｂａＩＨ片段克隆。

The chitinase gene of Heliothis armigera single nucleocapsid nucleopolyhedrovirus(HaSNPV) was localized on BamHI E, BglII E,EcoRI G,HindIII F,XbaI H,BamHI+HindIII M,and BamHI+XbaI H fragments, using the α P 32 dATP labelled recombinant plasmid harboring Choristoneura fumiferana multiple nucleocapsid nucleopolyhedrovirus (CfMNPV)DNA HindIII M which contains chitinase gene as the probe. The XbaI H fragment of HaSNPV was cloned into pTZ 19R vector for further sequencing the chitinase gene.