Molecular Cloning and Induced Expression Analysis of CD8 Gene from Humphead Snapper

doi:10.13560/j.cnki.biotech.bull.1985.2017-0095

Abstract

Abstract: This work aims to characterize CD8 gene of humphead snapper，Lutjanus sanguineus. The full length cDNA sequences of CD8α and CD8β genes were cloned by homologous cloning and rapid amplification of cDNA ends（RACE）from humphead snapper，and their tissue distribution and expression pattern after induction with immunostimulants were also analyzed by real time quantitative PCR（qRT-PCR）. The CD8α’s cDNA consisted of 1 576 bp encoding 225 amino acids，and the CD8β’s cDNA consisted of 1 486 bp encoding 210 amino acids. The predicted CD8α and CD8β proteins were similar in structure，both contained a signal peptide，immunoglobulin superfamily variable domain，hinge region，transmembrane domain and cytoplasmic tail. The qRT-PCR analysis showed that the highest level of CD8α and CD8β mRNA expression was found in thymus，followed by kidney，gill，intesine，skin，spleen，and head kidney. Furthermore，the mRNA levels of CD8α and CD8β in head kidney leucocytes was significantly up-regulated after 8 h treated with the stimulants lipopolysaccharide（LPS），concanavalin A（ConA），and polyinosinic-polycytidylic acid（PolyI∶C），respectively（P < 0.01）. And their expression levels in gill，anterior kidney，spleen，and intesine also increased at 24 h after stimulated with formalin-inactivated Vibrio harveyi.

Key words: Lutjanus sanguineus, CD8, gene cloning, induced expression

HUANG Yu-cong, LIANG Xiu-quan ,CAI Shuang-hu,LU Yi-shan,JIAN Ji-chang, WU Zao-he. Molecular Cloning and Induced Expression Analysis of CD8 Gene from Humphead Snapper[J]. Biotechnology Bulletin, 2017, 33(7): 169-178.

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