DNA Shuffling of Vitreoscilla Hemoglobin
- VernacularTitle:透明颤菌血红蛋白的DNA改组研究
- Author:
Ning YUAN
;
You-Jia HU
;
Chun-Bao ZHU
;
Bao-Quan ZHU
;
- Publication Type:Journal Article
- Keywords:
Vitreoscilla hemoglobin(VHb) Error-prone PCR DNA shuffling
- From:
China Biotechnology
2006;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
To improve the growth enhancement activity of Vitreoscilla hemoglobin(VHb), Vitreoscilla hemoglobin gene(vgb) was mutated by error-prone PCR and then reconstituted by DNA shuffling. The shuffling library was constructed by inserting the shuffled genes into the downstream of vgb natural promoter and transforming them into E.coli DH5?. Mutated active VHb proteins were first screened in test tubes according to host cell pellets color and then in shake flasks according to host pellets wet weight .One active mutant protein, VHb′042506, was obtained after second screening. It could increased the host wet weight by 31.25% and 58.75% than that of the control which bearing natural VHb under microaerobic and extremely microaerobic conditions, respectively. Sequencing and alignment results showed that 11 nucleotides were mutated, thus resulted in 4 amino acids changes occurred in this mutant protein. CO difference spectrum test also indicated that it had higher specific absorption.
