Sequence analysis and expression pattern ofSTARD3 cDNA in Chlamys farreri
In this paper,the sequence and expression characteristics of STARD3 in Chlamys farreri were studied to provide a basis for further comprehensive exploration of its gonadal development mechanism.STARD3 is a member of the steroidogenic acute regulatory protein related lipid transfer domain(START)family.This study using the existing transcriptome data of Chlamys farreri,obtained the CDS sequence of STARD3,analyzed the amino acid sequence of STARD3 using DNAMAN,compared its protein homology,and finally constructed the phylogenetic tree using neighbor joining(NJ)method.At the same time,RNA was extracted in different tissues(kidney,gill,mantle,adductor muscle,hepatopancreas,testis and ovary)and different stages(proliferation stage,growth stage and mature stage)of Chlamys farreri,and then was reverse transcribed into cDNA,as a template to detect the relative expression of the gene in different tissues and different stages.The results showed that the coding sequence(CDS)of the gene was 1 512 bp,encoding 503 amino acids.The amino acid sequence comparison of different species showed that the STARD3 of Chlamys farreri had high homology with other species,up to 92.01％,which included START domain,MLN64 Net alterminal(METAL)domain and diphenylalanine(FF)-acid orbital(AT)(FFAT)motif,as well as the amino acid residues required for biological functions,such as Arg,Ser,Phe,Leu and Tyr.Phylogenetic analysis showed that STARD3 protein sequence of Chlamys farreri first clustered with scallops such as Mizuhopecten yessoensis,followed by Mytilus edulis and Mytilus coruscus,next followed by Sepia pharaonis and Octopus bimaculoides belonging to the same mollusca phyla,then clustered with lower vertebrates,and finally with higher mammals,which proved that the phylogenetic relationship of STARD3 in Chlamys farreri was consistent with the taxonomic position of the species.The results of qRTet alPCR in kidney,hepatopancreas,mantle,adductor muscle,gill,ovary and testis showed that STARD3 was detected in the above organizations,indicating that the expression was extensive.In females,the expression level of STARD3 in ovary was the highest(P＜0.05),followed by that in gill and kidney.In males,the expression of STARD3 in testis was higher than that of kidney,and significantly higher than that of other tissues.To sum up,the expression of STARD3 gene in sexual gland was significantly higher than that in other tissues(P＜0.05),followed by the expression in kidney and gill,indicating that the gene mainly played a role in sexual gland and also played a certain role in other tissues.In the gonadal development cycle,taking the testis and ovarys of scallops at different stages as the research objects,the result showed that the expression in the testis during the proliferative stage was significantly higher than that in the other two stages,and the expression in this stage was 3.6 times higher than that in the growth stage and 13 times higher than that in the mature stage.In the ovary,the expression in proliferative stage was significantly higher than that in the growth stage,slightly higher than that in the mature stage,and showed a trend of first decreasing and then increasing with the cycle change.Therefore,this study preliminarily reveals that STARD3 plays a major role in early gonadal development by detecting the expression of STARD3 in various tissues and gonadal development cycle of Chlamys farreri,and also has certain functions in other tissues,but its function and specific mechanism of action need to be further studied.
万方数据
维普
