博莱霉素A_5的类酶性研究

CONSTRUCTION OF VACCINA VIRUS RECOMBINANT EXPRESSION 5aG STRAIN RABIES VIRUS GLYCOPROTEIN GENE AGAINST RABIES

研究了博莱霉素Ａ５（ＢＬＭＡ５）与ＤＮＡ作用前后的光谱性质及初步的动力学性质，发现ＢＬＭＡ５可催化ＤＮＡ氧化断链，而且符合米氏动力学．通过对ＢＬＭＡ５的催化效率、底物专一性以及温度、ｐＨ、金属离子对ＢＬＭＡ５催化ＤＮＡ断链反应影响的研究，发现ＢＬＭＡ５的作用行为在许多方面与酶的催化行为非常类似，提出ＢＬＭＡ５是一种小分子生物催化剂的看法．

The 5aG strain virus glycoprotein gene was inserted into the vaccinia virus transfection vector pJ120,which the coding sequence was controlled under the vaccinia virus late promoter P11.Plasmid DNA was introduced into vaccinia virus Tiantan strain by LIPOFECTIN method.Through the homologous recombination,this gene was integrated into TK region of vaccinia virus genome. Under BrdU selection pressure,one strain of recombinant virus has been picked out.Immunofluorecence test and Western Blotting showed that the lecombinant virus expressed 5aG strain rabies virus glycoprotein on the surface of infected cell.The molecular weight of glycoprotein approximated 64×10 3.Mice experiments showed that all the mices which immunized with recombinant virus acquired protection against lethaldose of CVS strain rabies virus challenge intracerebally.After mice was inocubated with recombinant virus, the titer of neutralizing antibody againt rabies virus was as high as 1:560 on the 14th day.