Expession of Microneme Protein 4 from Eimeria tenella in the Methylotrophic Yeast Pichia pastoris

Zhou Sai, An Jian, Zhang Peng, W. Lixia
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Abstract

Eimeria ten Ella is a major cause of coccidiosis in chickens, which can lead to serious economic impact on poultry production worldwide. Developing a cost effective sub-unit vaccine may be helpful to decrease loss in the poultry enterprise. Here, we describe expressing etMIC4, which is one of the microneme proteins. It is a 218 kDa microneme protein from Eimeria ten Ella. In this study, RNA was extracted from E. ten Ella sporulated oocysts and N-(etMIC4-N) and C terminal (etMIC4-C) genes were cloned by PCR, according to published etMIC4 cDNA sequence. Blast analysis showed that 773bp of 5 'end and 883bp of 3' end respectively shared more than 98% identy with etMIC4. At last, etMIC4-N was successfully expressed in pichia pastor is and identified by Western Blot, which set up a platform for further research.
柔嫩艾美耳球虫微基因蛋白4在毕赤酵母中的表达
艾美耳球虫是引起鸡球虫病的主要原因,可对世界范围内的家禽生产造成严重的经济影响。开发具有成本效益的亚单位疫苗可能有助于减少家禽企业的损失。在这里,我们描述了表达etMIC4,这是微素蛋白之一。它是一个218 kDa的微素蛋白,来自艾美耳球虫。本研究从E. ten Ella孢子卵囊中提取RNA,根据已发表的etMIC4 cDNA序列,采用PCR方法克隆N-(etMIC4-N)和C端(etMIC4-C)基因。Blast分析显示,5′端773bp和3′端883bp分别与etMIC4具有98%以上的同源性。最后,etMIC4-N在毕赤酵母中成功表达,并经Western Blot鉴定,为进一步研究奠定了基础。
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