运用酶切自连法分析鉴定肺炎链球菌体内诱导基因.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 20 10 5 庞丹 王虹 尹楠林 杨晓亮 胥文春 尹一兵 张雪梅 ( , 400016) : 肺炎链球菌是严重侵袭性感染 上呼吸道感染最重要的条件致病菌之一, 分析鉴定体内诱导基因序列变得尤 为重要本研究利用酶切自连法成功从 129 个体内诱导表达的肺炎链球菌中获得 13 个融合重组自杀质粒, 通过与肺炎链球 菌株 TIGR4基因组序列的同源性比对, 共得到 10 个不同的体内诱导基因, 其中8 个是从血液中筛选出的, 另 2 个为从肺组织 中筛选出; 通过分析得到 18 个开放阅读框, 其中大部分为已知功能基因, 参与细菌多种生命活动, 有 2 个为未知功能基因, 编 码假想蛋白可见, 酶切自连法可有效用于筛选基因的分析鉴定 : 酶切自连法 自杀质粒 肺炎链球菌 体内诱导基因 Analysis and Identification of in vivo Induced Genes of Strep tococcus p neumun iae byEnzyme SelfligationM ethod Pang Dan W ang Hong Y in Nanlin Yang X iaoliang X W ench n Y in Y ib ing Zhang X em ei (Key Laboratory of DiagnosticM edicineDesignated byM inistry of Education, ChongqingM edical University, Chongqing 400016) Abstrac:t Strep tococcuspneu oniae is one of themost mi portant opport n istic pathogenaswh ich can lead to serio s invasive infec tion and pper resp iratory tract infection. Th s, it becam e mi portant to analysis and identification of in vivo ind ced genes seq ences. In this st dy, the recomb inant s icide plasm idswere harvested by enzyme selfligation m ethod, and seq enced to obtain the in vivo ind ced genes seq ences, the f nctions ofwh ich w ere predicted by bioinformatics analysis. 13 recomb inant s icide plasm ids, from the 129 individ alsw ith in the ind cib le expression ofStrep tococcus pneu oniae, were harvested. A total of 10 d ifferent in vivo ind ced genesw ere ob tained by b lasting w ith strep tococcuspneu uniae TIGR4 genom e seq ence, 8 ofwh ich are screened o t from the blood, wh ile the other two from the l ng tiss esw ere screened. By analyzing the available 18 open read ing frames, m ost ofwh ich are known f nctional genes involved in a variety of life activities of bacteria. There were two genes of nknown f nction, encoded hypothetical proteins. The st dy proved that the enzyme selfl