CAJ | 학술논문

抗菌肽是蛙类非特异性免疫的重要组成部分,蛙类对环境变化比较敏感,但仍能分布于广泛的地理区域,抗菌肽的多样性和组成是揭示其适应机制的基础。本研究通过RT-PCR技术,从1只黑斑侧褶蛙的皮肤组织中克隆了9条不同的cDNA序列,分属5个家族：Esculentin-1、Esculentin-2、Temporin、Nigrocin-1和Nigrocin-2。其中Esculentin-1、Esculentin-2、Temporin和Nigrocin-1为阳离子抗菌肽,等电点（PI）8.72～9.63,分子量（Mw）1495.87～4862.9,预测的二级结构都具有α-螺旋结构。Nigrocin-2为新发现的一类酸性抗菌肽,等电点PI为4.0,Mw为1798.06,带1个负电荷,预测二级结构不具有α-螺旋结构,呈无规则卷曲。这9条抗菌肽cDNA来自50个阳性克隆,显示出黑斑侧褶蛙抗菌肽具多样性,特殊抗菌肽的发现也预示着黑斑侧褶蛙适应复杂多变环境、分布广泛的生理潜能,但具体功能还需进一步研究。
항균태시와류비특이성면역적중요조성부분,와류대배경변화비교민감,단잉능분포우엄범적지리구역,항균태적다양성화조성시게시기괄응궤제적기출。본연구통과RT-PCR기술,종1지흑반측습와적피부조직중극륭료9조불동적cDNA서렬,분속5개가족：Esculentin-1、Esculentin-2、Temporin、Nigrocin-1화Nigrocin-2。기중Esculentin-1、Esculentin-2、Temporin화Nigrocin-1위양리자항균태,등전점（PI）8.72～9.63,분자량（Mw）1495.87～4862.9,예측적이급결구도구유α-라선결구。Nigrocin-2위신발현적일류산성항균태,등전점PI위4.0,Mw위1798.06,대1개부전하,예측이급결구불구유α-라선결구,정무규칙권곡。저9조항균태cDNA래자50개양성극륭,현시출흑반측습와항균태구다양성,특수항균태적발현야예시착흑반측습와괄응복잡다변배경、분포엄범적생리잠능,단구체공능환수진일보연구。
Antimicrobial peptides（AMPs） are a major component of innate immunity of amphibians and play essential roles in amphibian survival and evolution.Frogs are sensitive to environmental changes but are widely dispersed across heterogeneous geographic regions.The composition and diversity of AMPs provide clues to their adaptive evolution.We identified 9 different AMP cDNAs from fresh skin tissue of a black spotted frog（Pelophylax nigromaculata） by using RT - PCR technology.These cDNAs were assigned to 5 families,four of which,namely Esculentin - 1,Esculentin - 2,Temporin and Nigrocin - 1,were all cationic peptides with molecular weights between 1495.87 and 4862.9,and theoretical isoelectric points（PI） between 8.72 and 9.63.All were predicted to form an a - helix in water solution.The fifth family was a novel acidic AMP first identified in amphibians,hereby named Nigrocin -2,with molecular weight 1798.06,1 net negative charge and theoretical PI of 4.0.The predicted secondary structure was a random coil in water solution.These 9 cDNAs were identified from 50 clones, demonstrating the diversity of AMPs in the black spotted frog.The identification of a special acidic AMP suggested the physiological potential of the frog to survive unusual environment factors across its large and heterogeneous range.However,the physiological function of the acidic AMP is yet to be studied.