韶关学院学报
韶關學院學報
소관학원학보
Journal of Shaoguan University(Social Science Edition)
2008年
9期
88~91
,共null页
细菌光敏色素 体内重组 诱导表达 优化
細菌光敏色素 體內重組 誘導錶達 優化
세균광민색소 체내중조 유도표체 우화
AphA(26-320)-PCB ; reconstitution in vivo ; inducment expression; expression optimization
将蓝藻PCC7120细菌光敏色素缺失突变体AphA(26-320)基因和血红素氧化酶(h01)及胆绿素还原(PcyA)的基因共同转化E.Coil BL21(DE3),在IPTG的诱导下实现体内共同表达,进行体内重组,生成了具有可逆光致变色活性的AphA(26-320)-PCB,对重组茵的培养条件,诱导条件进行优化,确定适合合成AphA(26-320)-PCB的培养时间,诱导温度,摇床振荡速度等。
將藍藻PCC7120細菌光敏色素缺失突變體AphA(26-320)基因和血紅素氧化酶(h01)及膽綠素還原(PcyA)的基因共同轉化E.Coil BL21(DE3),在IPTG的誘導下實現體內共同錶達,進行體內重組,生成瞭具有可逆光緻變色活性的AphA(26-320)-PCB,對重組茵的培養條件,誘導條件進行優化,確定適閤閤成AphA(26-320)-PCB的培養時間,誘導溫度,搖床振盪速度等。
장람조PCC7120세균광민색소결실돌변체AphA(26-320)기인화혈홍소양화매(h01)급담록소환원(PcyA)적기인공동전화E.Coil BL21(DE3),재IPTG적유도하실현체내공동표체,진행체내중조,생성료구유가역광치변색활성적AphA(26-320)-PCB,대중조인적배양조건,유도조건진행우화,학정괄합합성AphA(26-320)-PCB적배양시간,유도온도,요상진탕속도등。
The aphA (26-320) was introduced into E. Coli BL21 (DE3) together with genes hol (encoding heme oxygenase) and pcyA to realize the autoassembly of the Cyanobacterial phytochrome AphA(26-320) and the PCB. Upon induction,recombinant E. Coli used the cellular pool of heme to produce AphA (26-320)-PCB with spectroscopic properties similar to those of the same protein produced endogenously in cyanobacteria, AphA(26-320)- PCB was red/infrared (R/FR)photochromic. After studying on the cultural and inducement condition of recombi- nant AphA(26-320)-PCB,The experiment condition was optimized to determine the suitable inducement tempera- ture and time.