Expression of phenylalanine ammonia-lyase cDNA from rice in E. coli BL21DE3
- VernacularTitle:水稻苯丙氨酸氨解酶基因在大肠杆菌BL21 DE3中的表达
- Author:
Zhunan CAI
;
Yingnian YU
;
Jianhong LUO
;
Yuli QIAN
;
Xiangruo CHEN
- Publication Type:Journal Article
- Keywords:
Phenylalanine ammonia-lyase;
Gene rearrangement;
Plasmid;
Escherichia coli
- From:
Chinese Journal of Pathophysiology
1986;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To study the expression and its kinetics of rice phenylalanine ammonia-lyase gene encoding into E. coli as the basis of treatment for phenylketouria. METHODS: The phenylalanine ammonia-lyase-1-cDNA(rPAL-1-cDNA) from rice was recombined into E. coli high expression vector pET-28c and transformed into E. coli host strain BL21DE3. Engineering bacteria was then inducted by isopropyl-?-D-thiogalactoside (IPTG) for 1, 3, 5, 7 hours, in order to obtain high level expression. RESULTS: After induction, the expression level of fusion protein was 21.40%, 30.60%, 35.40%, 35.43% respectively. The fusion protein exhibited a band of 78 6 kD on SDS-PAGE analysis,but was not found in controls.The target protein was mainly existed in the form of inclusion body. CONCLUSION:Rice PAL gene expressing E. coli was established by gentic engineering technique.
