Zhan Bo, Gao Yuan-Yuan, Lin Wei-Ping, Jia Guang-Tao, Sun Tong-Yi, Lee Wen-Hui
School of Biological Science and Technology, Weifang Medical University, Weifang 261053, China.
School of Clinic Medicine, Weifang Medical University, Weifang 261053, China.
Zhongguo Zhong Yao Za Zhi. 2016 Feb;41(4):630-635. doi: 10.4268/cjcmm20160414.
The skin transcriptome of Bufu bufo gargarizans was determined by conventional methods. A novel full length cDNA coding for a Cathelicidin precursor was identified by transcriptomic data assembling, annotation and blast search of corresponding data banks. According to the known processing methods of Cathelicidin family members, present reported novel Cathelicidin precursor of B. bufo gargarizans might be cleaved at 2 possible sites of the same precursor and generate both BG-CATH25 and BG-CATH29 as mature molecules. The deduced BG-CATH25 and BG-CATH29 were synthesized with purity>95% to evaluate the properties and bactericidal activities. The secondary structural characteristics of both BG-CATH25 and BG-CATH29 in different solutions were determined by Circular Dichroism (CD) Analysis. CD results indicated that random coil conformation were the main structural elements for both BG-CATH25 and BG-CATH29 in different buffer systems. Antimicrobial activities against tested bacterial strains were carried out by plating method. Both BG-CATH25 and BG-CATH29 showed strong antibacterial activities against Aeromonas hydrophila, with MIC values of 1.25, 10 mg•L⁻¹, respectively. However, both of them showed weak bactericidal activities against human pathogenic bacteria, like Escherichia coli (ATCC25922),Staphylococcus aureus (ATCC25923)and Pseudomonas aeruginosa (ATCC 27853).
采用常规方法测定了中华大蟾蜍的皮肤转录组。通过转录组数据组装、注释以及对相应数据库的比对搜索,鉴定出一种编码Cathelicidin前体的新型全长cDNA。根据Cathelicidin家族成员已知的加工方式,目前报道的中华大蟾蜍新型Cathelicidin前体可能在同一前体的2个可能位点处被切割,生成BG - CATH25和BG - CATH29作为成熟分子。合成了推导的BG - CATH25和BG - CATH29,纯度>95%,以评估其性质和杀菌活性。通过圆二色性(CD)分析确定了BG - CATH25和BG - CATH29在不同溶液中的二级结构特征。CD结果表明,在不同缓冲体系中,无规卷曲构象是BG - CATH25和BG - CATH29的主要结构元件。采用平板法对受试细菌菌株进行抗菌活性测试。BG - CATH25和BG - CATH29对嗜水气单胞菌均表现出较强的抗菌活性,MIC值分别为1.25、10 mg•L⁻¹。然而,它们对人类病原菌,如大肠杆菌(ATCC25922)、金黄色葡萄球菌(ATCC25923)和铜绿假单胞菌(ATCC 27853)的杀菌活性较弱。