Noordhoek G T, Hermans P W, Paul A N, Schouls L M, van der Sluis J J, van Embden J D
Laboratory for Bacteriology, National Institute of Public Health and Environmental Hygiene, Bilthoven, The Netherlands.
Microb Pathog. 1989 Jan;6(1):29-42. doi: 10.1016/0882-4010(89)90005-3.
In an attempt to identify antigenic differences between Treponema pallidum subsp. pallidum (T. pallidum) and Treponema pallidum subsp. pertenue (T. pertenue) a gene bank of T. pertenue was constructed in lambda vector EMBL3. Clones carrying the T. pertenue gene encoding a 190 kDa protein, TyF1, were selected and the DNA was expressed in E. coli. TyF1 was shown to be closely related, but slightly different from the previously cloned T. pallidum antigen TpF1. TyF1 and TpF1 are high molecular weight antigens of about 190 kDa, which dissociate into 19 kDa subunits after heat treatment in presence of SDS. The difference between the two proteins is most obvious after treatment with proteinase K, which yields a 115 kDa component from TyF1 and a 95 kDa component from TpF1, as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The structural genes encoding TyF1 and TpF1 were sequenced and the predicted amino acid sequences differed in a single amino acid residue at position 40, which is arginine in TyF1 and glutamine in TpF1. Similarities TyF1 and TpF1 with the previously described 4D antigen are discussed. The antibody response to TyF1 and TpF1 seems higher in syphilis patients than in yaws patients. The possibility of using the difference between these T. pallidum and the T. Pertenue antigens for serological discrimination of syphilis and yaws is discussed.
为了鉴定梅毒螺旋体苍白亚种（T. pallidum）和雅司螺旋体亚种（T. pertenue）之间的抗原差异，构建了T. pertenue的λ噬菌体载体EMBL3基因文库。挑选出携带编码190 kDa蛋白TyF1的T. pertenue基因的克隆，并在大肠杆菌中表达该DNA。结果表明，TyF1与先前克隆的梅毒螺旋体抗原TpF1密切相关，但略有不同。TyF1和TpF1是约190 kDa的高分子量抗原，在SDS存在下热处理后可解离成19 kDa的亚基。用蛋白酶K处理后，两种蛋白质之间的差异最为明显，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）测定，TyF1产生115 kDa的组分，TpF1产生95 kDa的组分。对编码TyF1和TpF1的结构基因进行了测序，预测的氨基酸序列在第40位的单个氨基酸残基上有所不同，TyF1中的该残基为精氨酸，TpF1中的为谷氨酰胺。讨论了TyF1和TpF1与先前描述的4D抗原的相似性。梅毒患者对TyF1和TpF1的抗体反应似乎高于雅司病患者。讨论了利用这些梅毒螺旋体和雅司螺旋体抗原之间的差异进行梅毒和雅司病血清学鉴别的可能性。
