摘要
为了给小麦主要过敏原CM16蛋白的重组表达和免疫活性鉴定等研究奠定基础,通过生物信息学方法设计并合成简并性引物,利用RT-PCR技术对小麦主要过敏原CM16基因进行克隆,并进行序列分析。结果表明,克隆获得了小麦主要过敏原CM16基因。基因开放阅读框为432个碱基(包括终止密码子),编码143个氨基酸。该序列编码的蛋白相对分子质量约为15 782,等电点为5.17。序列同源性分析发现其与国外报道的已知小麦CM16基因具有很高的同源性(同源性为99%),因此认为其系小麦过敏原基因,在GenBank数据库中的登录号为EU883599。
In order to clone CM16 gene from wheat cultivar Chinese Spring （Triticum aestivum L）, CM16 gene sequences was compared and the conservative domains among these sequences were used for designing degenerate primers. The RT- PCR methods were applied to clone the full-length CM16 from Triticum aestivum L and the sequence was analyzed. Results showed that the major allergen gene CM16 of Triticurn aestivum L was cloned. The cloned fragment was with 432 bp open reading frame coding for 143 amino acids. Sequence analysis showed that the estimated molecular mass was 15 782 with pI 5.17. Sequence analysis showed that this gene shared high identities with CM16 previous reported in Triticum aestivum L. The deduced protein was therefore regarded as the major allergen of CM16 of Triticum aestivurn L（ Gen-Bank database entry No. EU883599）. CM16 gene cloned will be used for further expression and immunocharacterization of Triticum aestivum L and the diagnosis of allergies, immune therapy.
出处
《麦类作物学报》
CAS
CSCD
北大核心
2009年第1期9-12,共4页
Journal of Triticeae Crops
基金
广东省科技重点专项(2003A3080502)
深圳市科技计划项目(200326)
