Urdea M S, Merryweather J P, Mullenbach G T, Coit D, Heberlein U, Valenzuela P, Barr P J
Proc Natl Acad Sci U S A. 1983 Dec;80(24):7461-5. doi: 10.1073/pnas.80.24.7461.
We have chemically synthesized and expressed in yeast a gene coding for human epidermal growth factor (urogastrone), a 53-amino-acid polypeptide that has been shown to promote epithelial cell proliferation and to inhibit gastric acid secretion. The synthetic gene, consisting of 170 base pairs, was designed with yeast-preferred codons and assembled by enzymatic ligation of synthetic fragments produced by phosphoramidite chemistry. The DNA synthesis protocol used allows for facile synthesis of oligonucleotides larger than 50 bases. Yeast cells were transformed with plasmids containing the synthetic gene under control of a yeast glyceraldehyde-3-phosphate dehydrogenase gene promoter and were shown to synthesize a biologically active human epidermal growth factor.
我们已经通过化学合成并在酵母中表达了一个编码人表皮生长因子（尿抑胃素）的基因，尿抑胃素是一种含53个氨基酸的多肽，已被证明可促进上皮细胞增殖并抑制胃酸分泌。该合成基因由170个碱基对组成，采用酵母偏好密码子设计，并通过亚磷酰胺化学法产生的合成片段进行酶促连接组装而成。所使用的DNA合成方案能够轻松合成大于50个碱基的寡核苷酸。用含有该合成基因的质粒转化酵母细胞，该基因受酵母甘油醛-3-磷酸脱氢酶基因启动子控制，结果表明酵母细胞能够合成具有生物活性的人表皮生长因子。
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