Mullenbach G T, Tabrizi A, Blacher R W, Steimer K S
J Biol Chem. 1986 Jan 15;261(2):719-22.
A synthetic gene encoding the platelet-derived factor, connective tissue activating peptide-III (CTAP-III) (Castor, C.W., Miller, J.W., and Walz, D.A. (1983) Proc. Natl. Acad. Sci. U. S. A. 80, 765-769), has been expressed and secreted from Saccharomyces cerevisiae by using a yeast expression vector and an alpha-factor leader segment. Mitogenic activity reported for naturally derived CTAP-III has been demonstrated here for recombinant CTAP-III. Active CTAP-III has been purified to apparent homogeneity. Structural studies have confirmed its identity. A general approach for the facile synthesis of genes is presented which has permitted the assembly of the entire structural gene and flanking regions (280 base pairs) from 20 oligomers in a single annealing and ligation reaction pool.
通过使用酵母表达载体和α-因子前导序列,已在酿酒酵母中表达并分泌了编码血小板衍生因子结缔组织活化肽III(CTAP-III)的合成基因(卡斯特,C.W.,米勒,J.W.,和瓦尔兹,D.A.(1983年)《美国国家科学院院刊》80,765 - 769)。本文已证明重组CTAP-III具有天然来源的CTAP-III所报道的促有丝分裂活性。活性CTAP-III已被纯化至表观均一性。结构研究已证实其身份。本文介绍了一种简便的基因合成通用方法,该方法允许在单个退火和连接反应池中由20个寡聚物组装整个结构基因及其侧翼区域(280个碱基对)。