研究了分离特定的基因片段，通过这些大肠杆菌DH5α菌株产生大肠杆菌素的利用。在第一阶段的大肠杆菌DH5α菌株时，他们被迫与质粒pcolx作出反应，这是在LB培养基上生长在37C到OD 280为1。然后用Mitcomycin C诱导大肠杆菌素生产菌株。观察未来6小时用SDS-聚丙烯酰胺凝胶电泳技术的细胞。收集的数据，通过物理观测的凝胶。研究结果如下
车道1标记的数据进行14.4,18.4,25,35,45,60.2，96K。车道2车道3车道未纯化的大肠菌素4-9小时诱导培养标本。这里的大肠杆菌质粒pcolx DH5alpha细胞转化。观察细胞生长6小时。样品被取均匀小时。此外，COL X基因的表达在大肠杆菌细胞内bl21ai阿拉伯糖诱导时OD值达到0.5。在2小时内，收获的细胞开始其中通过超声破碎细胞，离心。可溶性蛋白进一步从游离细胞中提取纯化，通过亲和层析与协议的帮助。
Research was conducted to isolate the particular gene fragments that were utilized by these E.Coli DH5 strain to produce Colicin. In the first stage the E.Coli DH5 strain when they were made to react with plasmid pColX, which was grown in LB media at 37C until the OD 280 was 1.0. They were then treated with Mitcomycin C to induce the colicin production of the strain. The cells were observed for the next 6 hours by using the SDS-PAGE technique. The data was collected by physically observing the gel. The results were as follows.
The Lane 1 markers data were 14.4,18.4,25,35,45,60.2, 96k. The Lane 2 purified colicin Ia Lane 3 uninduced Lanes 4-9 hourly samples of induced culture. Here the E coli DH5alpha cells transformed with the plasmid pColX. Cell growth was observed for a period of 6 hours. Samples were taken even hour. Further, the gene of Col X was expressed in terms of BL21AI cells of E coli within the induction of arabinose when 0.5 as OD was reached. Over a 2 hour period, harvesting the cells began wherein through sonication the cells were broken in order to be centrifugated. Soluble proteins were further extracted from free cells for purification through affinity of nickel chromatography with the help of protocols.
In addition, the pColX and the pColE1 consisted of higher sequences similar to bp 496 ranging from one gene to other with the help of a protein named lysis for starting the colicin activity gene codon (Braun et al 1994). This was initially done because the western blot made use of antibodies of anti Colla showing the formation of a band wherein colX involved primers of sequencing for the Colla terminal ends.