摘要
以野生细菌F0 30 3的总DNA为模板 ,采用细菌 16SrDNA的通用引物 ,通过PCR的方法扩增到一条约 1.5Kb的 16SrDNA片段。连接到pGEM -T -easy克隆载体上 ,并用化学法转化E .coliDH5α。用EcoRI对 5个随机挑取的转化子进行的酶切分析表明这 5个转化子皆为阳性。DNA测序表明该PCR扩增到的 16SrDNA片段长为 14 75核苷酸。与GenBank上已提交的16SrDNA进行的比对 (BLAST)表明野生细菌F0 30 3归属于沙雷氏属。由VectorNTISuite 6软件构建的系统发育树表明与深红色沙雷氏细菌 (Serratiarubidaea)亲源关系最近 ,在核苷酸水平有 97.8%的相似性。
A 1.5Kb of 16SrDNA fragment amplified through gen eral PCR with the template of field bacterium F0303 total DNA,and bacteria 16SrDNA universal primers,was ligated into pGEM-Teasy vector which is classic cloning vector for PCR products,and then was taken to transform chemically competent cell of E.coli DH5α.Restriction enzyme analysis of the 5 randomly selected transformants,mediated by EcoR I,demonstrates they were all positive.The BLAST of the 1475 bp of 16SrDNA sequence obtained by DNA sequencing,with all 16SrDNA sequences in GenBank indicated that the field bacterium F0303 belongs to Serratia genus and the phylogenic tree of the field bacterium F0303,which was generated by Vector NTI Suite 6 software,basing on 16SrDNA full-length sequence,showed that it is most close to Serratia rubidaea with 97.8% of identity.
出处
《生物信息学》
2005年第1期1-4,18,共5页
Chinese Journal of Bioinformatics
