水产学报2017，Vol.41Issue(11)：1687-1698,12.DOI:10.11964/jfc.20161110618
马氏珠母贝TLR6基因的克隆、序列分析与表达
Cloning, sequence analysis and expression studies on PmTLR6 gene from Pinctada fucata martensii
摘要
Abstract
TLR6 (Toll like receptor 6) is a kind of pattern recognition receptors and plays an important role in resisting microorganism infection.To study the function of TLR6 in the immune response of P.fucata martensii, in this study, a full length ofPm-TLR6 was obtained using rapid amplification of cDNA ends (RACE) technology from P.fucata martensii.The expression patterns ofPm-TLR6 in all tissues and its sequential expression in the hemolymph after Vibrio harveyi stimulation and nucleus insertion operation were further detected by Quantitative Real-Time PCR technology.Results showed that the total length ofPm-TLR6 cDNA was 2295 bp, including a 5'UTR of 94 bp, a 3' UTR of 89 bp and an open reading frame (ORF) of 2112 bp which encodes 703 amino acids.Multiple sequence alignment indicated that TLR6 was highly conservative among species.The protein encoded by Pm-TLR6 has a transmembrane domain, several leucine rich repeats(LRRs) and a TIR domain, conforming to the characteristics of TLRs family.qRT-PCR data revealed thatPm-TLR6 was expressed in all tested tissues, including hepatopancreas, hemocytes, gill, gonads, adductor muscle and mantle, with the highest expression in hepatopancreas (P<0.05).AfterVibrio harveyi injection, the expression level ofPm-TLR6 increased at 2 h (9 fold vs.control), then dropped to normal levels at 6 h and began to increase at 16 h, with the highest level of expression appearing at 24 h (29.4 fold vs.control,p<0.05).The result of nucleus insertion surgery showed thatPm-TLR6 expression level was not significantly changed at 5 d and 10 d, and began to increase at 15 d and 20 d (P>0.05).Its expression reached the maximum level at 30 d with significant difference (5 fold vs.control,p<0.05).These results indicated thatPm-TLR6 may play an important role in the immune defense reaction of P.fucata martensii.关键词
马氏珠母贝/TLR6/基因克隆/表达分析Key words
Pinctada fucata martensii/TLR6/gene cloning/expression analysis分类
生物科学引用本文复制引用
吴羽媛,郭志颖,梁海鹰,林丽旋,郝瑞娟,焦钰..马氏珠母贝TLR6基因的克隆、序列分析与表达[J].水产学报,2017,41(11):1687-1698,12.基金项目
国家自然科学基金(31472306) （31472306）
广东省科技计划项目(2012A031100010) （2012A031100010）
广东省海港建设与渔业产业发展专项(A201608B15) （A201608B15）
国家级大学生创新创业训练项目(201410566005) （201410566005）
广东海洋大学大学生创新创业训练项目(CXXL2014012) National Natural Science Foundation of China (31472306) （CXXL2014012）
Science and Technology Plan Projects of Guangdong Province, China (2012A031100010) （2012A031100010）
Harbor Construction and Development of Special Fisheries Industry of Guangdong Province, China (A201608B15) （A201608B15）
National College Students' Innovation Entrepreneurship Training Program (201410566005) （201410566005）
College Students' Innovation Entrepreneurship Training Projects of Guangdong Ocean University (CXXL2014012) （CXXL2014012）
