长爪沙鼠β-防御素(defβ-83)基因的克隆与鉴定

Clone and Identification of β-defensin(defβ-83) Gene in the Mongolian Gerbil

目的克隆长爪沙鼠β-防御素基因序列,并对其进行鉴定及分析。方法从长爪沙鼠小肠中提取总RNA,根据GeneBank中大小鼠的β-防御素的基因序列,通过防御素基因的保守性设计引物,采用RT-PCR技术得到预期的PCR产物,将所得的片段进行克隆、测序,并应用相关生物信息学软件对序列进行鉴定和分析,序列提交genbank。结果Blast比较发现最终测序的结果与小鼠β-防御素-1和大鼠β-防御素-1的同源性全都大于80%,genebank登录号为EU834052。结论经测序鉴定证实该PCR产物为长爪沙鼠β-防御素基因1的一部分,本研究为深入探讨长爪沙鼠β-防御素的生物学活性奠定基础。

Objective To Clone β-defensin gene in Mongolian gerbil. Methods The primers was designed according to the published sequences in GenBank. A RT-PCR method was applied to clone the defensin beta 1 gene in small intestine of the Mongolian gerbil. The PCR products was sequenced and blasted with related sequences. Results The sequenced results presented that the product was highly identical to the defensin beta 1 gene in rat and mouse. The identification between the present sequence and rat or mouse was over 80%, the GenBank Accession Number is EU834052. Conclusions The acquired sequence was a portion of defensin beta 1 gene, which presented highly identical to the same gene in rat and mouse. The further step field of the experimental Gerbils β-defensin is the biological activity.