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目的探讨蛋白表达技术在支原体研究中的作用。方法以解脲脲原体（ｕｒｅａｐｌａｓｍａｕｒｅａｌｙｔｉｃｕｍＵＵ）为例，根据所研究ＵＵ的目的ＤＮＡ片段设计一对特异性引物，经ＰＣＲ扩增得到目的ＤＮＡ，克隆后运用表达质粒载体ｐＧＥＸ２Ｔ在ＩＰＴＧ诱导下成功诱导了蛋白的表达，并用Ｗｅｓｔｅｒｎ印迹法检测其抗原活性。结果所表达的蛋白具有很好的抗原活性。结论蛋白表达应用于支原体研究中，为研究支原体的致病性与种类或型别的关系提供了重要的手段。实验中我们体会到：（１）在设计引物时，要注意避开ＵＧＡ，可用ＵＧＧ代替。（２）ＩＰＴＧ的浓度从０．１ｍｍｏｌ／Ｌ～１ｍｍｏｌ／Ｌ均有报道，我们实验证实两种浓度在诱导蛋白表达中无差别，因而推荐使用０．１ｍｍｏｌ／Ｌ。（３）蛋白诱导的时间以４～４．５ｈ为最佳。
Objective To investigate the role of protein expression in the study of Mycoplasma. Methods Taking Ureaplasma urealyticumUU as an example, a pair of specific primers were designed according to the target DNA fragment of UU. The target DNA was amplified by PCR. After cloning, the plasmid pGEX2T was successfully induced by IPTG The protein expression was induced and its antigenic activity was detected by Western blotting. Results The expressed protein has good antigenic activity. Conclusion The protein expression is applied to mycoplasma research, which provides an important means for studying the relationship between pathogenicity and species or genotype of mycoplasma. Experiment we experience: (1) in the design of primers, should pay attention to avoid UGA, available UGG instead. (2) The concentration of IPTG is reported from 0.1 mmol / L to 1 mmol / L. Our experiments confirmed that there is no difference between the two concentrations in inducing protein expression, therefore 0.1 mmol / L is recommended. (3) protein-induced time to 4 ~ 4.5h for the best.