CAJ | 학술논문

目的：为获得PGEX-5X-I载体与EGFR-COM112的融合蛋白。方法将EGFR-COM112基因克隆入已构建好的载体PGEX-5X-I中,转化大肠杆菌Top10。该菌株经IPTG诱导后,高效表达出融合蛋白PGEX-5X-I-EGFR-COM112,经PCR和测序鉴定正确的重组质粒。经PBS稀释复性、Ni离子亲和层析柱纯化,获得目的蛋白。结果 SDS-PAGE分析表明,在38 kD左右有一条明显的特异性蛋白条带。结论成功地制备了EGFR-COM112的载体并纯化其蛋白表达。
목적：위획득PGEX-5X-I재체여EGFR-COM112적융합단백。방법장EGFR-COM112기인극륭입이구건호적재체PGEX-5X-I중,전화대장간균Top10。해균주경IPTG유도후,고효표체출융합단백PGEX-5X-I-EGFR-COM112,경PCR화측서감정정학적중조질립。경PBS희석복성、Ni리자친화층석주순화,획득목적단백。결과 SDS-PAGE분석표명,재38 kD좌우유일조명현적특이성단백조대。결론성공지제비료EGFR-COM112적재체병순화기단백표체。
Objective In order to obtain PGEX -5x -I car ier and EGFR - COM112 fusion protein. Methods EGFR - COM112 gene cloning into car ier has built PGEX-5X-I. Transformation of escherichia coli Top10 may need. Top 10 after IPTG induction, ef icient expressed fusion protein PGEX-5X-I-EGFR-COM112, via PCR and sequencing appraisal right recombinant plasmid. The PBS dilution method, Ni ion af inity chromatography column purification, get the target protein. Results Sds-page analysis showed that there is a obvious specificity around 38 kD protein bands. Conclusion The car ier of the EGFR-COM112 successful y prepared and purified protein expression.