文献类型：期刊文献

英文题名：Chromosomal integration of the Vitreoscilla hemoglobin gene and its physiological actions in Tremella fuciformis

作者：Zhu, Hu[1]; Wang, Tian-Wen[1]; Sun, Shu-Jing[2]; Shen, Ya-Ling[1]; Wei, Dong-Zhi[1]

机构：[1] State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China; [2] Mycological Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China

年份：2006

卷号：72

期号：4

起止页码：770

外文期刊名：Applied Microbiology and Biotechnology

收录：EI(收录号：20064010144621)

语种：英文

外文关键词：Assays - Cells - Chromosomes - DNA - Enzymes - Escherichia coli - Fermentation - Hemoglobin - Oxygen - Reaction kinetics

摘要：The Vitreoscilla hemoglobin (VHb) gene was expressed in yeast-like conidia (YLCs) of Tremella fuciformis (T. fuciformis) to increase cell density in submerged fermentation by enhancing oxygen uptake. With the intention of doing this, an integrated expression vector containing the VHb gene and the hygromycin B phosphotransferase (hph) gene derived from Escherichia coli (E. coli) as the selectable marker was constructed, and then transformed into protoplasts of YLCs from T. fuciformis with restriction enzyme-mediated DNA integration (REMI). Hygromycin-resistant transformants had been generated during the transformation. Molecular evidences including PCR assay, Southern blotting, and Western blot analysis indicated the VHb gene had been integrated into the genome of transgenic T. fuciformis strains and was expressed successfully. Shake-flask fermentation and bioreactor cultivation results showed that the expression of VHb in this fungus could enhance growth of YLCs. The final cell density was higher in the culture of VHb-expressing strain than that of the wild-type strain. Moreover, these results also suggested that CaMV35S promoter was capable of driving the expression of heterologous genes in T. fuciformis. ? 2006 Springer-Verlag.