水产学报2018，Vol.42Issue(5)：646-652,7.DOI:10.11964/jfc.20170110689
日本沼虾N-乙酰-β-D-氨基葡萄糖苷酶基因克隆及KK-42对其表达的影响
Molecular cloning of N-acetyl-β-D-glucosaminidase(NAGase)gene and the effect of KK-42 on NAGase gene in Macrobrachium nipponense
摘要
Abstract
In order to further explore the molecular mechanism that KK-42 shortens the molt cycle of the juvenile prawn Macrobrachium nipponense,the full-length cDNA sequence of NAGase,the rate-limiting enzyme in chitin catabolism,was cloned from carapace by RACE technique.The relative expression of NAGase mRNA and its activity in cuticular tissue were determined before and after KK-42 treatment. Sequence analysis showed that the full length NAGase cDNA was 2 536 bp,encoding 617 amino acids.Homology analysis indicated that NAGase was less conserved and had the highest similarity of 68% to that from Litopenaeus vannamei.Phylogenetic analys-is showed that the amino acid sequence of NAGase from M.nipponense was clustered into one major group with that from Portunus trituberculatus,L.vannamei,and Fenneropenaeus chinensis.The NAGase sequence from L. vannamei and F.chinensis shared more similarities with each other,M.nipponense belonged to a separate branch. The mRNA concentration of cuticular NAGase in control group peaked at premolt D0stage.Once treated by KK-42, the mRNA content increased significantly, with a 253% rise in D4phase at 3 h, as well as a 226% or 187% rise in C or D0phase at 6 h. The activity of NAGase rose gradually from C to D4. KK-42 treatment could cause signi-ficant increase of NAGase activity in C and D0stages, especially in C stage, during which the activity increased by 11.26, 5.99 and 7.15 folds, respectively, at 3, 6 and 12 h. The results above suggest that the induction effect of KK-42 on the cuticular NAGase of M.nipponense may be one of the molecular mechanisms to shorten the molt cycle.关键词
日本沼虾/N-乙酰-β-D-氨基葡萄糖苷酶/克隆/KK-42Key words
Macrobrachium nipponense/N-acetyl-β-D-glucosaminidase/clone/KK-42分类
生物科学引用本文复制引用
吕艳杰,关建义,杜娟,宁黔冀..日本沼虾N-乙酰-β-D-氨基葡萄糖苷酶基因克隆及KK-42对其表达的影响[J].水产学报,2018,42(5):646-652,7.基金项目
河南省教育厅科学技术研究重点项目(14A240003) （14A240003）
河南师范大学青年科学基金(2015QK15) （2015QK15）
天津市动植物抗性重点实验室开放基金(5201049160124) （5201049160124）
Basic Key Project of Science and Technology Research of Henan Province Educational Com-mission(14A240003) （14A240003）
Youth Science Foundation of Henan Normal University(2015QK15) （2015QK15）
Open Fund of Tianjin Key Laboratory of Animal and Plant Resistance(5201049160124) （5201049160124）
