利用抑制性差减杂交技术(Suppression subtractive hybridization,SSH)对虾夷扇贝闭壳肌积累类胡萝卜素相关基因进行筛查和分析。以虾夷扇贝橘红色闭壳肌和白色闭壳肌cDNA构建正反向差减文库,结合454测序技术在正向差减文库中测序得到2 640条序列,拼接得到50个contigs(重叠群);反向差减文库中测序得到2 496条序列,拼接得到187个contigs。经过同源性比对分析,共获得150多个差异表达基因,从中选择3种清道夫受体SRB1、SRF2和SRCR作为候选基因进行研究,对其在虾夷扇贝各组织中的表达量水平进行检测,初步阐述了其在虾夷扇贝中的组织表达规律。根据结果推测在虾夷扇贝橘红色闭壳肌中,清道夫受体有可能是调节类胡萝卜素累积和转运的关键基因。本研究为虾夷扇贝闭壳肌类胡萝卜素累积相关通路的探索提供了初步研究的基础数据。
Differentially expressed genes relating to carotenoids accumulation in adductor muscle of Yesso scallop(Patinopecten yessoensis) were isolated using suppression subtractive hybridization(SSH) technique.The cDNA of orange and white adductor muscle was used to construct the forward and reverse subtracted libraries.The cDNA fragments differentially expressed in both subtracted libraries were sequenced using 454 sequencing technique.In the forward subtracted library,50 contigs were obtained from 2640 sequences;in the reverse subtracted library,187 contigs were obtained from 2496 sequences.There were more than 150 genes differentially expressed from which three scavenger receptor genes such as SRB1,SRF2 and SRCR were chosen as candidate genes.The analysis of quantitative expression of scavenger receptor genes in all tissues of Yesso scallop was carried out.The results revealed that the scavenger receptors may play a key role in mediating the cellular uptake of carotenoids in Yesso scallops.The results provided preliminary foundation for exploring the pathway of carotenoids accumulation.