Rogers D T, Lemire J M, Bostian K A
Proc Natl Acad Sci U S A. 1982 Apr;79(7):2157-61. doi: 10.1073/pnas.79.7.2157.
Two clones from a lambda phage collection containing yeast genes regulated by inorganic phosphate were shown by low-stringency hybridization to select three mRNAs that direct the in vitro synthesis of repressible acid phosphatase (EC 3.1.3.2) polypeptides p60, p58, and p56. By higher stringency hybridization one yeast fragment [8 kilobases (kb)] selects p60 mRNA and the other (5 kb) selects p56 mRNA. These EcoRI digestion fragments were subcloned in yeast transformation vectors and hybridization selection assignments were confirmed by measuring enzyme and mRNA levels in transformants. Enzyme and mRNA levels in (8-kb) high copy number transformants grown in high inorganic phosphate medium revealed a hitherto undetected acid phosphatase protein, P57, which is believed to correspond to the constitutive enzyme encoded by PHO3. The identify of the 8-kb fragment purported to contain the PHO5/PHO3 genes was confirmed by genetic mapping of an integrated copy of this fragment. The site of integration of the 5-kb fragment was demonstrated to be unlinked to the PHO5/PHO3 genes.
通过低严谨度杂交显示，从一个含有受无机磷酸盐调控的酵母基因的λ噬菌体文库中筛选出的两个克隆，能选择出三种信使核糖核酸（mRNA），它们指导体外合成可阻遏酸性磷酸酶（EC 3.1.3.2）的多肽p60、p58和p56。通过更高严谨度杂交，一个酵母片段[8千碱基（kb）]选择p60 mRNA，另一个（5 kb）选择p56 mRNA。这些EcoRI消化片段被亚克隆到酵母转化载体中，通过测量转化体中的酶和mRNA水平，证实了杂交选择的结果。在高无机磷酸盐培养基中生长的（8 kb）高拷贝数转化体中的酶和mRNA水平显示出一种迄今未检测到的酸性磷酸酶蛋白P57，据信它对应于由PHO3编码的组成型酶。通过对该片段的整合拷贝进行遗传定位，证实了据称含有PHO5/PHO3基因的8 kb片段的身份。已证明5 kb片段的整合位点与PHO5/PHO3基因不连锁。
