栉江珧wnt4基因cDNA的克隆表达及调控
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中国海洋大学 海水养殖教育部重点实验室, 山东 青岛 266003

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王昌勃(1988-),男,博士.E-mail:c-b-wang@163.com

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S917

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国家海洋公益性行业科研专项(201305005);国家科技支撑计划项目(2011BAD13B01);教育部博士点基金(20130132110009).


Molecular cloning and expression analysis of wnt4 from Atrina pectinata and effects of estradiol-17β on its expression
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Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao 266003, China

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    摘要:

    本研究运用同源克隆技术和RACE技术克隆了栉江珧(基因cDNA全长序列。该基因序列全长为1493 bp,其中开放阅读框1074 bp,编码由357个氨基酸组成的蛋白。氨基酸序列分析表明,栉江珧Paracentrotus lividus)等物种具有高度的同源性。荧光定量PCR分析表明,基因表达量与性腺成熟度相关,并且整个繁殖周期卵巢表达量均显著高于精巢,说明基因主要参与了栉江珧的早期胚胎发育。在胚胎发育早期(囊胚期和原肠期)基因可能在栉江珧早期发育阶段参与了某些器官的形成。17<0.05);短时间处理,17<0.05)。

    Abstract:

    The pen shell is a large, fan-shaped bivalve that is widely distributed in the Indian and western Pacific oceans. However, little is known about the molecular regulation of its gonad development. The (wingless-type MMTV integration site family, member 4) gene, a member of the family, encodes a growth factor that plays an important role in gonad differentiation and development. In this study, the complete cDNA sequence of ovary by homology-based cloning and rapid amplification of cDNA ends techniques. The full-length cDNA sequence of was 1493 bp long and included an open reading frame of 1074 bp encoding a 357-amino acid protein with a predicted molecular weight of 39.5 kD and a theoretical isoelectric point of 8.94. Multiple alignment and phylogenetic analysis of wnt proteins showed that the wnt4 sequence contained conserved domains of the wnt family. In the phylogenetic analysis, the Chlamys farreri and lower similarity to the wnts of echinoderms, amphibians, and mammals. Quantitative real-time polymerase chain reaction analysis revealed that transcripts were present at higher levels in the female mantle and gonad of wnt4 transcripts was related to gonad maturity, and the transcription of was significantly ovary-predominant during the entire reproductive cycle. These patterns of transcription suggested that A. pectinata. During the early larval development stage of was detected in early embryogenesis (morula stage and gastrula stage), when it was 500 times that in adults, and then the transcript level fell sharply during the trochophore and D-shaped stages. Therefore, wnt4 may be involved in the formation of certain organs in the stages of early larval development. When the sex steroid estradiol-17 through feedback regulation ( treatment significantly induced transcription of , regulated by estradiol-17, is involved in a variety of biological processes in different tissues, especially during early embryogenesis and gonad development. Further research is required to verify whether the transcript level of can be used as an index of testis development.

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王昌勃,李琪,于瑞海,孔令锋,于红.栉江珧wnt4基因cDNA的克隆表达及调控[J].中国水产科学,2016,23(5):1041-1051
WANG Changbo, LI Qi, YU Ruihai, KONG Lingfeng, YU Hong. Molecular cloning and expression analysis of wnt4 from Atrina pectinata and effects of estradiol-17β on its expression[J]. Journal of Fishery Sciences of China,2016,23(5):1041-1051

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  • 在线发布日期: 2016-09-05
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