Cloning and Expression Analysis of IKKε Homologous Gene from Pinctada martensii
Fisheries College, Guangdong Ocean University, Zhanjiang, 524025, China
International Journal of Marine Science, 2014, Vol. 4, No. 56 doi: 10.5376/ijms.2014.04.0056
Received: 12 Jul., 2018 Accepted: 28 Jul., 2018 Published: 12 Oct., 2014
© 2014 BioPublisher Publishing Platform
This article was first published in Genomics and Applied Biology (2014，33(2): 266-272) in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Jiao Y., Du X.D., Wang Q.H., Huang R.L., and Deng Y.W., 2014, Cloning and expression analysis of IKKε homologous gene from Pinctada martensii, International Journal of Marine Science, 4(56): 1-6 (doi: 10.5376/ijms.2014.04.0056)
IKKε (inhibitor of NF-κB kinases ε) is a key member in the NF-κB (nucleur factor κB) signal pathway and participates in the regulation of the cell differentiation, development, apoptosis and immune response. In this study, primers were designed based on the unigene sequence annotated as IKKε in the transcriptome database of Pinctada martensii, using rapid amplification of cDNA ends (RACE) technology, the full length sequence of IKKε gene was obtained from Pinctada martensii (PmIKKε). At the same time, the mRNA expression of PmIKKε in 6 tissues of Pinctada martensii was detected by Real-time PCR technology. The results showed that the obtained full length of PmIKKε gene cDNA was 2,843 bp, among which 5' UTR was 116 bp and 3' UTR was 516 bp, containing 27 bp polyA tail. The open reading frame (ORF) was 2,211 bp encoding 737 amino acid residues. The predicted molecular weight was 85.07 kD, isoelectric point was 6.17. Multiple sequence alignment indicated that PmIKKε had highly homology among other species IKKε and had 45% sequence identity with Crassostrea gigas. The software analysis results showed that the N-terminal region contained a protein kinase functional domain and an activation site of mitogen-activated protein kinase (MAPK), The C-terminal region contained a leucine zipper (LZ) domain and a helix-loop-helix (HLH) motif. Real-time PCR analysis demonstrated that PmIKKε was constitutively expressed in six studied tissues (hepatopancreas, gonad, hemolymph, gill, mantle, aductor muscle) in P. martensii, and the highest expression was found in hepatopancreas. This study could provide a theoretical basis for the further study of the function of PmIKKε in growth, development and innate immunity in of Pinctada martensii.
IKKε; Pinctada martensii ; Gene clone; Real-time PCR