Martin M C, Schneider D, Bruton C J, Chater K F, Hardisson C
Area de Microbiología, Facultad de Medicina and Instituto Universitario de Biotecnología de Asturias, Universidad de Oviedo, Spain.
J Bacteriol. 1997 Dec;179(24):7784-9. doi: 10.1128/jb.179.24.7784-7789.1997.
By using a PCR approach based on conserved regions of ADP-glucose pyrophosphorylases, a glgC gene was cloned from Streptomyces coelicolor A3(2). The deduced glgC gene product showed end-to-end relatedness to other bacterial ADP-glucose pyrophosphorylases. The glgC gene is about 1,000 kb from the leftmost chromosome end and is not closely linked to either of the two glgB genes of S. coelicolor, which encode glycogen branching enzymes active in different locations in differentiated colonies. Disruption of glgC eliminated only the first of two temporal peaks of ADP-glucose pyrophosphorylase activity and glycogen accumulation and prevented cytologically observable glycogen accumulation in the substrate mycelium of colonies (phase I), while glycogen deposition in young spore chains (phase II) remained readily detectable. The cloned glgC gene therefore encodes an ADP-glucose pyrophosphorylase essential only for phase I (and it is therefore named glgCI). A second, phase II-specific, glgC gene should also exist in S. coelicolor, though it was not detected by hybridization analysis.
通过使用基于ADP - 葡萄糖焦磷酸化酶保守区域的PCR方法,从天蓝色链霉菌A3(2)中克隆出一个glgC基因。推导的glgC基因产物与其他细菌的ADP - 葡萄糖焦磷酸化酶显示出端对端的相关性。glgC基因距离最左边的染色体末端约1000 kb,并且与天蓝色链霉菌的两个glgB基因均不紧密连锁,这两个glgB基因编码在分化菌落的不同位置具有活性的糖原分支酶。glgC的破坏仅消除了ADP - 葡萄糖焦磷酸化酶活性和糖原积累的两个时间峰中的第一个,并阻止了菌落底物菌丝体(I期)中细胞学上可观察到的糖原积累,而年轻孢子链(II期)中的糖原沉积仍易于检测到。因此,克隆的glgC基因编码仅对I期必不可少的ADP - 葡萄糖焦磷酸化酶(因此将其命名为glgCI)。天蓝色链霉菌中应该也存在第二个II期特异性的glgC基因,尽管通过杂交分析未检测到。