Production of synthetic spider dragline silk protein in Pichia pastoris
Production of synthetic spider dragline silk protein in Pichia pastoris
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DOI：
10.1007/s002530050884
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发表时间：
1997-01-01
影响因子：
5
通讯作者：
Bedzyk, LA
中科院分区：
文献类型：
--
作者：
Fahnestock, SR;Bedzyk, LA
甲醇营养酵母巴斯德毕赤酵母被测试为生产长的、重复的蛋白质聚合物的宿主。在甲醇诱导的AOX1启动子的控制下，蜘蛛拖丝蛋白的设计类似物的合成基因很容易得到高水平的表达。含有多个基因拷贝的转化体产生了高水平的丝蛋白，但由于转化时的基因重排，其大小发生了各种变化。长达3000个密码子或更长的基因可以表达，但没有证据表明类似基因在大肠杆菌中普遍存在截断合成，尽管超过1600个密码子的基因表达效率低于较短的基因。产丝的巴氏假单胞菌菌株在没有经过选择的情况下至少可以加倍100次。
The methylotrophic yeast Pichia pastoris was tested as a host for the production of long, repetitive protein polymers. Synthetic genes for a designed analog of a spider dragline silk protein were readily expressed at high levels under control of the methanol-inducible AOX1 promoter. Transformants containing multiple gene copies produced elevated levels of silk protein, but of a variety of altered sizes as a result of gene rearrangements at the time of transformation. Genes up to 3000 codons in length or longer could be expressed with no evidence of the prevalent truncated synthesis observed for similar genes in Escherichia coli, though genes longer than 1600 codons were expressed less efficiently than shorter genes. Silk-producing P. pastoris strains were stable without selection for at least 100 doublings.