In order to provided clues to the development of Japonicin?pb, we cloned the Japonicin?pb from Paa boulengeri and investigated its expression patterns among different growth temperatures and tissues. The com?plete open reading frame ( ORF) sequence of Japonicin?pb precursor was amplified by RT?PCR according to the transcriptome database of Paa boulengeri constructed by our laboratory. The sequential structure of Japoni?cin?pb was analyzed by the method of domain alignment. The expression patterns of Japonicin?pb among differ?ent growth temperatures (15, 18, 21, 24, 27 and 30 ℃) and tissues (blood, muscle, liver and skin) were analyzed by real?time PCR. The results showed that according to the domain alignment, 3 Japonicin?pb precur?sors had the same N terminal signal peptide, middle spacer region, but the mature peptides were 19, 20 and 12 amino acid residues, named Japonicin?1apb, Japonicin?1bpb and Japonicin?3pb, respectively. The expression patterns showed that Japonicin?1apb was mainly expressed in skin, and its expression abundance increased gradually from 15 to 30 ℃;the expression level of Japonicin?1bpb among different tissues and growth temper?atures were no significant difference; Japonicin?pb mainly distributed in liver, and it was not sensitive to the response of growth temperature. These results provide important clues for the functional study and the develop?ment of Japonicin?pb.%本试验旨在克隆棘腹蛙来源Japonicin?pb,了解该抗菌肽在不同生长温度及组织下的表达规律,为Japonicin?pb的开发提供线索. 基于本实验室构建的棘腹蛙皮肤转录组数据库,利用反转录PCR从棘腹蛙皮肤组织中克隆Japonicin?pb前体序列,通过结构域比对的方法分析其结构. 利用Real?time PCR检测3种Japonicin?pb在不同生长温度( 15、18、21、24、27和30℃)及组织(血液、肌肉、肝脏和皮肤)的表达图谱. 结果表明,本试验克隆到3种Japonicin?pb前体序列,结构域比对表明,三者拥有相同的N端信号肽和中间间隔区,长度分别为19、20和12个氨基酸残基的肽序列,命名为Japonicin?1apb、Japonicin?1bpb和Japonicin?3pb. Japonicin?1apb主要在皮肤组织中表达,且表达量随着生长温度的升高而逐渐增加;Japonicin?1bpb在不同组织或生长温度下的表达量差异均不明显;Japonicin?3pb主要在肝脏中表达,且生长温度对其影响不大. 这些结果为棘腹蛙源3 种Japonicin?pb后续功能研究以及开发应用提供了重要的序列及表达谱信息.
展开▼
