Campbell L A, Kuo C C, Thissen R W, Grayston J T
Department of Pathobiology, University of Washington, Seattle 98195.
Infect Immun. 1989 Jan;57(1):71-5. doi: 10.1128/iai.57.1.71-75.1989.
Chlamydia sp. strain TWAR is a unique Chlamydia sp. that causes acute respiratory disease. A gene bank consisting of TWAR isolate AR-39 DNA in pUC19 was screened with anti-AR-39 rabbit immune sera. Two positive clones were isolated that contained 7.3-kilobase (pLC1) and 14.9-kilobase (pLC2) plasmids. Restriction mapping and hybridization studies showed that both pLC1 and pLC2 contained a common 4.2-kilobase PstI fragment. Plasmids were used as templates of in vitro transcription-translation. All three plasmids had a novel protein product of ca. 75 kilodaltons not found in the vector alone. Western blots showed that this protein reacted with anti-TWAR rabbit immune sera and with human immune serum from an individual who had proven TWAR infection. Whole-cell lysates of TWAR demonstrated a protein having the same molecular weight and immunoreactivity as the recombinant gene product. This protein was also recognized by rabbit immune serum against Chlamydia psittaci or Chlamydia trachomatis. Southern hybridizations with the cloned fragment as a probe of digests of other Chlamydia spp. showed weakly hybridizing fragments. These results suggest that we have isolated a gene encoding a protein recognized during human TWAR infection that contains some sequences shared among Chlamydia spp.
沙眼衣原体TWAR株是一种独特的沙眼衣原体，可引起急性呼吸道疾病。用抗AR-39兔免疫血清筛选由pUC19中的TWAR分离株AR-39 DNA组成的基因文库。分离出两个阳性克隆，分别含有7.3千碱基（pLC1）和14.9千碱基（pLC2）的质粒。限制性图谱分析和杂交研究表明，pLC1和pLC2都含有一个共同的4.2千碱基PstI片段。质粒用作体外转录-翻译的模板。所有三种质粒都有一种约75千道尔顿的新蛋白质产物，单独在载体中未发现。蛋白质印迹法表明，这种蛋白质与抗TWAR兔免疫血清以及来自已证实感染TWAR的个体的人免疫血清发生反应。TWAR的全细胞裂解物显示出一种与重组基因产物具有相同分子量和免疫反应性的蛋白质。这种蛋白质也被抗鹦鹉热衣原体或沙眼衣原体的兔免疫血清识别。用克隆片段作为探针与其他沙眼衣原体属的消化物进行Southern杂交，显示出弱杂交片段。这些结果表明，我们分离出了一个编码在人类TWAR感染期间被识别的蛋白质的基因，该蛋白质含有沙眼衣原体属之间共享的一些序列。
