Bailey M J, Thomas C M, Cockayne A, Strugnell R A, Penn C W
School of Biological Sciences, University of Birmingham, UK.
J Gen Microbiol. 1989 Sep;135(9):2365-78. doi: 10.1099/00221287-135-9-2365.
A library of Treponema pallidum genomic DNA fragments produced by partial Sau3A digestion was established in Escherichia coli K12 using the plasmid vector pAT153. The library was screened using immune syphilitic rabbit serum and six recombinant phenotypes expressing eight treponemal polypeptides were detected. With two exceptions, all the recombinant gene products were the same size as polypeptides detected on Western immunoblots of T. pallidum. The genes encoding three novel gene products, with molecular masses in SDS-PAGE of 42, 17 and 15.5 kDa, which had not been cloned previously from T. pallidum were also identified. Monoclonal antibodies which reacted with four of the eight recombinant polypeptides were generated.
使用质粒载体pAT153，在大肠杆菌K12中建立了由部分Sau3A消化产生的梅毒螺旋体基因组DNA片段文库。用免疫梅毒兔血清筛选该文库，检测到表达8种梅毒螺旋体多肽的6种重组表型。除两个例外，所有重组基因产物的大小与梅毒螺旋体Western免疫印迹上检测到的多肽相同。还鉴定了编码三种新基因产物的基因，其在SDS-PAGE中的分子量分别为42、17和15.5 kDa，这些基因以前未从梅毒螺旋体中克隆出来。产生了与8种重组多肽中的4种发生反应的单克隆抗体。
