对虾病毒HHNBV DNA构建质粒的研究与分析

用ＰＵＣ１８构建了对虾病毒ＨＨＮＢＶＤＮＡ重组质粒，提取后经斑点杂交及酶切分析，证实质粒中插入片段为病毒ＤＮＡ，其中０５＃质粒的插入片段大于２Ｋｂ，与质粒的分子量相近。对０５＃质粒酶切后的Ｓｏｕｔｈｅｒｎ杂交亦取得了满意的结果。同时，还对０５＃质粒的插入片段进行了内切酶图谱分析，共进行了ＥｃｏＲｌ、Ｈｉｎｄｌｌｌ、Ｓｍａｌｌ、Ｐｓｔｌ、ＰＵＶｌ、Ｈｉｎｄｌｌ６种限制性内切酶分析，结果表明，只有Ｐｓｔｌ在插入片段的一端有一个酶切位点，酶切位点的确切位置有待于进一步确定。

THE CLONE AND STUDY OF THE HHNBV DNA FRAGMENT

The recombination clones of HHNBV was constracted using PUC18 Plasmid.The recombination plasmids was analysed with dot blot hybridization and restriction.The result showed that the foreign DNA fragments were parts of HHNBV DNA. The foreignfragments of 05 # plasmid was larger than 2kb, similar to the PUC18 plasmid. The southern blot hybridization of the 05 # plasmid was also successful.The restriction map of 05 # plasmid has been constructed.In six enzymes; EcoRl,Hindlll,Small, Pstl,PVUI and Hindll,only Pstl has a restriction site near one end.The relative postition of this restriction site was not determined yet.