【酸舔糖胜热甘露糖普cDNA克隆及部分特性的测】王姝.pdf

【酸舔糖胜热甘露糖普cDNA克隆及部分特性的测】王姝.pdf

摘 要 UDP-N-acetylglucosamine:β-D-mannosideβ-1N-acetylglucosaminyltransferasel(GnTl)可催化 将N-acetylglucosamine以β1-4键连接到N一糖链三 角型核区的β-[inked甘露糖上。我们从牛血浆中提纯了 该酶的底物,并使之用于酶的纯化。精制后的GnTIⅢI电泳 表现出一条主要带及一条较小的带,分子量分别为62kDa 及52kDa。根据该酶的胰酶水解片段氨基酸序列,设计引 物作PCR扩增,以扩增产物为探针,使用大鼠肾cDNA文库,通过斑点杂交法筛选GnTⅢIcDNA.并作序列分析。lesorleclnaliorcalactitrailiveriissues etibited vely litile activity wkile aigklerels were foued ir Al-e6 hepatoma and rat kidney(l2)These results agreedwiththe previous observation thatthebisecting GLcNACwaspresentinr-glutamyltranspeptidasespurifiedfromAB-66 hepatoma cell(l6)and from rat kidneycevelopmertalslages as foliow:3 wtcks.sweeks,9 weeks:10weeks weeksweek 21weeks.Preparationofthesubsirates: Fluorescence-labeled sugar wereobtained from bovine plasma by means ofpronase digestion, hydrazinolysis,.-r,andthea pyridylamination(PA-tion)according to Hase et al.