水产学报2023，Vol.47Issue(7)：27-37,11.DOI:10.11964/jfc.20220213319
马氏珠母贝丝裂原活化蛋白激酶p38的克隆与表达
Cloning and expression analysis of mitogen-activated protein kinase(MAPK)p38 in pearl oyster Pinctada fucata martensii
摘要
Abstract
The mitogen-activated protein kinase(MAPK)signaling pathway is crucial in cellular response to extra-cellular stimuli.This pathway utilizes serine/threonine-protein kinases that transmit extracellular signals through a phosphorylation cascade to cells.Rapid-amplification of cDNA ends(RACE)was utilized for cloning and quantit-ative PCR(qPCR)used for expression profiling of p38 MAPK in this study.Our findings reveal that the Pinctada fucata martensii(PmMAPK p38)has a full-length cDNA of 1 516 bp,an open reading frame(ORF)of 1 071 bp,and has an estimated molecular mass of 40.88 ku which is encoded 356 amino acids.Domain prediction analysis indicates that PmMAPK p38 has the typical MAPK family S_TKc domain and sequence alignment,tree construc-tion,and MatGAT calculation demonstrate its high similarity and conservation to MAPK genes in other species.Our qPCR results show that PmMAPK p38 is extensively expressed in various P.fucata martensii tissues,with the highest levels in hepatopancreas,followed by mantle,and the lowest levels in adductor muscle.Stimulation with LPS resulted in relative expression peaking at 2 h,decreasing to the least at 12 h.The greatest expression was roughly 5 times higher than the lowest.After stimulation with Vibrio harveyi,relative expression peaked at 2 h,decreased to the lowest at 8 h,with the highest expression approximately 4 times greater than the lowest.Our find-ings suggest that PmMAPK p38 may be a crucial component of the immune response in P.fucata martensii,and this study provides essential data for further investigation on the immune defense system of shellfish.关键词
马氏珠母贝/MAPK p38/免疫/基因克隆Key words
Pinctada fucata martensii/mitogen-activated protein kinase(MAPK)p38/immunity/gene cloning分类
生物科学引用本文复制引用
涂淏天,房晓宸,梁海鹰,雷倩楠,刘德凡..马氏珠母贝丝裂原活化蛋白激酶p38的克隆与表达[J].水产学报,2023,47(7):27-37,11.基金项目
National Natural Science Foundation of China(31472306) （31472306）
Natural Science Foundation of Guangdong Province(2023A1515012924,2021A1515010962) （2023A1515012924,2021A1515010962）
Science and Technology Special Fund of Guang-dong Province(2021A05250) （2021A05250）
Special Fund for Harbor Construction and Fishery Industry Development of Guang-dong Province(A201608B15) （A201608B15）
Sustainable Development Project of Shenzhen Science and Technology Program(KCXFZ20211020165547010)国家自然科学基金(31472306) （KCXFZ20211020165547010）
广东省自然科学基金(2023A1515012924,2021A1515010962) （2023A1515012924,2021A1515010962）
广东省科技专项(2021A05250) （2021A05250）
广东省海港建设与渔业产业发展专项(A201608B15) （A201608B15）
深圳市可持续发展专项(KCXFZ20211020165547010) （KCXFZ20211020165547010）
