【摘要】 从粘质沙雷氏菌(Serratia marcescensATCC14041)中克隆出几丁质酶基因(ChiC),将回收纯化的PCR产物与载体pMD18-T连接,构建成的重组质粒命名为pMD-Ch iC,将重组质粒转化到受体E.coliDH5α中进行克隆,经BamHI和NheI双酶切验证、核酸序列测定证实,重组质粒pMD-Ch iC含有几丁质酶C基因(ChiC)。利用生物信息学的方法,推测该粘质沙雷氏菌ChiC基因编码的蛋白质由480个氨基酸组成。预测该蛋白的等电点为5.63,分子量约为52kD。针对粘质沙雷氏菌中的几个几丁质酶基因做了进化树,进而验证了粘质沙雷氏菌(Serratia marcescens)几丁质酶(ChiC)在沙雷氏菌几丁质酶中的分类;同时对粘质沙雷氏菌几丁质酶C(ChiC)蛋白的高级结构作出了预测,得到其编码的属于18家族的蛋白质高级结构图谱。更多还原

【Abstract】 Chitinase gene ChiC from Serratia macescens ATCC14041 was cloned.The PCR product was inserted into the pMD18-T Vector(named pMD-ChiC).The recombinant plasmid was transformed into E.coli DH5α.By # sequenced and cleaved with Bam HI/Nhe I,it shows that recombinant plasmid contains ChiC.Bioinformatics analysis shows that the length of complete coding sequence was 1 443 bp,and the protein of the ChiC had a 480-residue precursor.The deduced isoelectric point was 5.63,and the molecular weight was about 52 kD.Chitinase cladogram of Serratia sp.was made,and the classification of Chitinase C in Serratia sp.was validated.Meanwhile the tertiary structure sketch map of the chitinase ChiC from S.marcescens was forecasted,and the senior structure sketch of the protein which belongs to 18 family was made.更多还原