将一株近海温泉耐热菌ZH1的16S rDNA克隆至pUCm-T载体进行测序,测序结果在NCB I上进行BLAST分析,并构建了系统发育树,将该菌株归属为Geobacillus sp.ZH1.在65℃条件下的产酶定性实验结果表明,该菌株产脂肪酶、木聚糖酶和碱性磷酸酶.以菌株ZH1的基因组DNA为模板,使用羧酸酯酶简并引物进行PCR扩增,将目的基因克隆至pUCm-T载体后进行测序,得到了741 bp的DNA片段.BLAST分析显示,目的基因预测编码羧酸酯酶,包含246个氨基酸.
The 16S rDNA from an inshore hot spring thermophilic strain ZH1 was cloned into pUCm-T vector and then sequenced.By analyzing the sequence against GenBank database with BLAST and constructing the phylogenetic tree,this strain was assigned to Geobacillus sp.ZH1.Qualitative analysis of its enzyme production at 65 ℃ showed that this strain could produce lipase,xylanase and alkaline phosphatase.To clone the carboxylesterase gene,it was amplified with the bacterial genomic DNA using a pair of degenerate primers.The target gene was cloned into pUCm-T vector and then sequenced.The obtained 741 bp DNA fragment was predicted to encode a carboxylesterase containing 246 amino acid residues with BLAST.