Berry A M, Paton J C, Glare E M, Hansman D, Catcheside D E
Department of Microbiology, Adelaide Children's Hospital, North Adelaide, South Australia.
Gene. 1988 Nov 30;71(2):299-305. doi: 10.1016/0378-1119(88)90046-7.
A gene bank of Sau3AI-generated Streptococcus pneumoniae DNA fragments was constructed in Escherichia coli K-12 by cloning into the BamHI site of the cosmid vector pHC79. One clone capable of cleaving the fluorogenic neuraminidase substrate 2'-(4-methylumbelliferyl)-alpha-D-N-acetyl-neuraminic acid was isolated. This activity was inhibited by treatment with a mouse antiserum raised against purified pneumococcal neuraminidase. The recombinant plasmid purified from this clone (designated pJCP301) contained approximately 3.0 kb of pneumococcal DNA. Western-blot analysis indicated that E. coli K-12[pJCP301] produced a 98-kDa polypeptide which reacted with antineuraminidase serum.
通过克隆到黏粒载体pHC79的BamHI位点，在大肠杆菌K-12中构建了由Sau3AI产生的肺炎链球菌DNA片段的基因文库。分离出一个能够切割荧光神经氨酸酶底物2'-(4-甲基伞形基)-α-D-N-乙酰神经氨酸的克隆。用针对纯化的肺炎球菌神经氨酸酶产生的小鼠抗血清处理可抑制这种活性。从该克隆中纯化的重组质粒（命名为pJCP301）含有约3.0 kb的肺炎球菌DNA。蛋白质印迹分析表明，大肠杆菌K-12[pJCP301]产生了一种与抗神经氨酸酶血清反应的98-kDa多肽。
