Yang Huajun, Liu Cheng, Zou Shaolan, Ma Yuanyuan, Hong Jiefang, Zhang Minhua
School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China,
Biotechnol Lett. 2014 Jun;36(6):1279-85. doi: 10.1007/s10529-014-1471-z. Epub 2014 Feb 22.
Introducing large numbers of target genes into the chromosome of Saccharomyces cerevisiae via δ-sequence-mediated integration is a good strategy for exploring the effects of gene dosage on expression and secretion of heterologous proteins. The expression of exogenous genes might be further improved through meiosis in an isogenic triploid. Here, a stable strain A-8 was screened from 35 sexual spore colonies obtained from an isogenic triploid integratively expressing bgl1 from Aspergillus aculeatus. The corresponding β-glucosidase activity in this strain was increased by ~120 % compared with the parent strain BGL-a. Measurement of doubling time, flow cytometry, and mating experiments further confirmed that A-8 was a spore-forming strain obtained from a triploid parent. Thus, combining δ-integration and meiosis in an isogenic triploid is a promising approach for improving the expression of exogenous proteins in S. cerevisiae.
通过δ序列介导的整合将大量目标基因导入酿酒酵母染色体是探索基因剂量对异源蛋白表达和分泌影响的良好策略。通过同基因三倍体中的减数分裂可能会进一步提高外源基因的表达。在此,从同基因三倍体中获得的35个性孢子菌落中筛选出了稳定菌株A-8,该同基因三倍体整合表达来自棘孢曲霉的bgl1。与亲本菌株BGL-a相比,该菌株中相应的β-葡萄糖苷酶活性提高了约120%。对倍增时间的测量、流式细胞术和交配实验进一步证实A-8是从三倍体亲本获得的产孢菌株。因此,在同基因三倍体中结合δ整合和减数分裂是提高酿酒酵母中外源蛋白表达的一种有前景的方法。