DNA shuffling of Vitreoscilla Hemoglobin

China Biotechnology

研究报告

DNA shuffling of Vitreoscilla Hemoglobin

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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 downstream of the vgb natural promoter and transforming them into E.coli DH5α. 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 can 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.

Key words： error-prone PCR DNA shuffling Vitreoscilla hemoglobin(VHb)

Received: 25 August 2006 Published: 25 November 2006

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Cite this article:

. DNA shuffling of Vitreoscilla Hemoglobin. China Biotechnology, 2006, 26(11): 14-19.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I11/14

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