Cloning and expression of breeding-& development-related gene Allatostatin-A receptor from Litopenaeus vannamei

Abstract:

Allatostatin-A receptor gene was cloned from Litopenaeus vannamei by RACE PCR, and the sequence length is 2 650 bp, including 1 425 bp of ORF, 1 170 bp of 5'-UTR, 55 bp of 3'-UTR, encoding 474 amino acids, forming an unstable hydrophilic protein with seven transmembrane regions, and subcellular localization is mainly located on the cytoplasmic membrane. The results of homology and phylogenetic analysis showed that the Allatostatin-A receptor gene of Litopenaeus vannamei has the highest homology with Penaeus monodon and Homarus americanus, and has the closest genetic relationship.The result of multiple amino acid sequence alignment showed that the C-terminal Arg after Allatostatin-A receptor amino acid TM7 is highly conserved among different species.Real time PCR results showed that the Allatostatin-A receptor gene was expressed in multiple tissues (eyestalk, brain, stomach, hepatopancreas, gill, gonad, intestine, muscle) of female and male Litopenaeus vannamei.Except for brain tissue, the relative expression of Allatostatin-A receptor gene in female Litopenaeus vannamei is higher than that in males.With the development of Litopenaeus vannamei larvae, the relative expression of Allatostatin-A receptor gene increased.The expression level of Allatostatin-A receptor gene was the highest in the hybrid offspring of imported male shrimp and Ecuadorian wild female shrimp. It is speculated that the Allatostatin-A receptor gene may be involved in the reproduction process of Litopenaeus vannamei,and whether it is involved in the growth regulation of Litopenaeus vannamei remains to be further studied.