论文部分内容阅读
目前认为肺炎衣原体(Chlamydia pneumonia,Cpn)除导致呼吸道疾病外,也是与冠心病相关的重要病原体。作为一种细胞内寄生的病原菌,Cpn激活宿主细胞信号通路,维护其在细胞内生长代谢,并导致疾病。肺炎衣原体基因Cpn0148可编码真核细胞样的丝/苏氨酸蛋白激酶,利用PCR技术扩增全长Cpn0148ORF,将其定向插入pGEX-6p原核表达载体,在大肠杆菌XL-1blue中表达,测序显示Cpn0148ORF全长1860bp,编码619个氨基酸,分子量大约70kD,将Cpn0148-GST融合蛋白免疫BALB/c小鼠获得抗Cpn0148多克隆抗体,间接免疫荧光法检测显示Cpn0148表达在菌体上。Cpn0148蛋白质的获得为进一步研究Cpn与宿主细胞的相互作用奠定了基础。
Chlamydia pneumonia (Cpn) is currently believed to be an important causative agent of coronary heart disease in addition to causing respiratory diseases. As an intracellular parasitic pathogen, Cpn activates host cell signaling pathways, sustaining its intracellular growth and metabolism, and leading to disease. Cpn0148 of Chlamydia pneumoniae can encode eukaryotic cell-like silk / threonine protein kinase. The full-length Cpn0148 ORF was amplified by PCR and inserted into pGEX-6p prokaryotic expression vector and expressed in E. coli XL-1blue. Sequencing showed Cpn0148 ORF is 1860bp in length and encodes 619 amino acids with a molecular weight of about 70kD. Cpn0148-GST fusion protein was immunized with BALB / c mouse to obtain anti-Cpn0148 polyclonal antibody. Cpn0148 expression was detected by indirect immunofluorescence assay. The obtainment of Cpn0148 protein lays the foundation for further study on the interaction between Cpn and host cells.