Sequence analysis of the chitinase gene cht1 of Bacillus circulans showed that it contained 2151 nucleotides,which codes the precursor of chitinase CHT1 with 717 amino acid residues.The nucleotide and deduced amino ac...Sequence analysis of the chitinase gene cht1 of Bacillus circulans showed that it contained 2151 nucleotides,which codes the precursor of chitinase CHT1 with 717 amino acid residues.The nucleotide and deduced amino acid sequences of cht1 showed 81% and 95% homology with those of ChiA of B.circulans WL-12,respectively.The cht1 gene was cloned into the Escherichia coli-Bacillus subtilis shuttle vector pSUGV4 and two recombinant plasmids, named pUSCH1 and pUSCH2 which contained 2.9kb and 4.0 kb insert respectively,were obtained.The recombinant plasmids were transformed into B.subtilis DB104 and WB600.Chitinase activity was detected both in transformed E.coli and B.subtilis.The DB104/pUSCH1 strain was found to be effective in the bio-controlling the infection of Magnaporthe grisea under greenhouse condition,which showed 71.67% decrease in rice disease incidence.展开更多
将含人巨细胞病毒(HCMV)DNA大片段的重组cos质粒转化入大肠杆菌DH5α。采用碱裂解法、煮沸法和SDS法提取纯化该大DNA重组cos质粒,并进行比较。通过限制性内切酶酶切分析,以HCMV DNA EcoRⅠ-Y片段作为探针,进行DNA印迹鉴定,并与33例19天...将含人巨细胞病毒(HCMV)DNA大片段的重组cos质粒转化入大肠杆菌DH5α。采用碱裂解法、煮沸法和SDS法提取纯化该大DNA重组cos质粒,并进行比较。通过限制性内切酶酶切分析,以HCMV DNA EcoRⅠ-Y片段作为探针,进行DNA印迹鉴定,并与33例19天～3月婴儿肝炎综合征患儿外周血白细胞DNA进行斑点杂交。结果提示:重组cos质粒转化效率为5.9×10~5转化子/μg DNA,碱裂解法可用于大质粒DNA提取,产量和纯度高,并省时。EcoRⅠ-Y片段与人白细胞DNA及其它疱疹病毒DNA无同源性,适宜予早期诊断HCMV感染,具有较血清学诊断及病毒分离培养特异性高、敏感性强和快速等优点。展开更多
文摘Sequence analysis of the chitinase gene cht1 of Bacillus circulans showed that it contained 2151 nucleotides,which codes the precursor of chitinase CHT1 with 717 amino acid residues.The nucleotide and deduced amino acid sequences of cht1 showed 81% and 95% homology with those of ChiA of B.circulans WL-12,respectively.The cht1 gene was cloned into the Escherichia coli-Bacillus subtilis shuttle vector pSUGV4 and two recombinant plasmids, named pUSCH1 and pUSCH2 which contained 2.9kb and 4.0 kb insert respectively,were obtained.The recombinant plasmids were transformed into B.subtilis DB104 and WB600.Chitinase activity was detected both in transformed E.coli and B.subtilis.The DB104/pUSCH1 strain was found to be effective in the bio-controlling the infection of Magnaporthe grisea under greenhouse condition,which showed 71.67% decrease in rice disease incidence.
文摘将含人巨细胞病毒(HCMV)DNA大片段的重组cos质粒转化入大肠杆菌DH5α。采用碱裂解法、煮沸法和SDS法提取纯化该大DNA重组cos质粒,并进行比较。通过限制性内切酶酶切分析,以HCMV DNA EcoRⅠ-Y片段作为探针,进行DNA印迹鉴定,并与33例19天～3月婴儿肝炎综合征患儿外周血白细胞DNA进行斑点杂交。结果提示:重组cos质粒转化效率为5.9×10~5转化子/μg DNA,碱裂解法可用于大质粒DNA提取,产量和纯度高,并省时。EcoRⅠ-Y片段与人白细胞DNA及其它疱疹病毒DNA无同源性,适宜予早期诊断HCMV感染,具有较血清学诊断及病毒分离培养特异性高、敏感性强和快速等优点。
