一种与抗生素调控基因afsk高度同源的新基因——Spy1的克隆

Cloning of Spy1,a Novel Homolog of afsk Gene

普那霉素是由始旋链霉菌产生的一种链阳性菌素类抗生素.目前国外对普那霉素的基因工程研究甚少,国内尚属空白,这阻碍了利用基因工程的方法来提高普那霉素产量的发展.本文通过对普那霉素产生菌——始旋链霉菌柯斯基因组文库的构建和菌落原位杂交,筛选出了与普那霉素高产密切相关的新基因所在的柯斯质粒,并通过酶切分析和Southern杂交确定了该基因所在的酶切片段,对酶切片段进行亚克隆测序,获得了与天蓝色链霉菌中抗生素调控基因afsk同源新基因的全长克隆,该新基因包含有1个2 127 bp的开放阅读框(ORF),编码708个氨基酸的蛋白,命名为Spy1.对该新基因的生物信息学初步分析表明,该基因编码的是丝/苏氨酸蛋白激酶.

Pristinamycin is an antibiotic of the streptogramin family that is produced by Streptomyces pristinaespiralis.Bacterial strains that engineered to produce pristinamycin producing were rare and non existed in China.We have constructed a cosmid library of a pristinamycin-producing strain and screened the colonies by in situ hybridization,in which a pristinamycin restriction fragment was used for Southern blotting.The target gene in full-length was successfully subcloned and named as Spy1.The construct contained a 2 127 bp open reading frame（ORF）,which encodes a protein of serine/threonine protein kinase of 708 amino acids.