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Differential expression and characterization analysis of a new gene with WD domains in fish oogenesis
WEN Jianjun; XIE Jing; LIU Shigui; GUI Jianfang; Gui, JF, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2001
Source PublicationSCIENCE IN CHINA SERIES C-LIFE SCIENCES
Volume44Issue:5Pages:541-553
AbstractA new gene with WD domains is cloned and characterized according to its differential transcription and expression between previtellogenic oocytes (phase I oocytes) and fully-grown oocytes (phase V oocytes) from natural gynogenetic silver crucian carp (Carassius auratus gibelio) by using the combinative methods of suppressive subtraction hybridization, SMART cDNA synthesis and RACE-PCR. The full-length cDNA is 1870 bp. Its 5 ' untranslated region is 210 bp, followed by an open reading frame of 990 bp, which has the typical vertebrate initiator codon of ANNATG. The open reading frame encodes a protein with 329 amino acids. It has 670 bp of 3 ' untranslated region and an AATAAA polyadenylation signal. Because it has 92% homology to STRAP (serine-threonine kinase receptor-associated protein), a recently reported gene, we named it FSTRAP (fish STRAP). Virtual Northern blotting indicated that the FSTRAP was transcribed in fully-grown oocytes (phase V oocytes), but not in previtellogenic oocytes (phase I oocytes). RT-PCR analysis showed that FSTRAP was transcribed in brain, heart, kidney, muscle, ovary, spleen and testis, but not in liver. And its mRNA could be detected in the oocytes from phase II to phase V. Western blotting also showed that FSTRAP protein could be detected in brain, heart, kidney, muscle, ovary, spleen and testis except liver. Results of Western blotting on various oocytes were also similar to the RT-PCR data. FSTRAP protein was not expressed in the previtellogenic oocytes. Its expression initiated from phase II oocytes after vitellogenesis, and was consistent with the mRNA transcription.
KeywordSilver Crucian Carp Oocyte Oogenesis Suppressive Subtractive Hybridization Wd Domain
DepartmentChinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; Sichuan Univ, Coll Life Sci, Chengdu 610064, Peoples R China
Subject AreaBiology
Indexed BySCI
WOS IDWOS:000171966600011
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/9920
Collection期刊论文
Corresponding AuthorGui, JF, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
WEN Jianjun,XIE Jing,LIU Shigui,et al. Differential expression and characterization analysis of a new gene with WD domains in fish oogenesis[J]. SCIENCE IN CHINA SERIES C-LIFE SCIENCES,2001,44(5):541-553.
APA WEN Jianjun,XIE Jing,LIU Shigui,GUI Jianfang,&Gui, JF, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.(2001).Differential expression and characterization analysis of a new gene with WD domains in fish oogenesis.SCIENCE IN CHINA SERIES C-LIFE SCIENCES,44(5),541-553.
MLA WEN Jianjun,et al."Differential expression and characterization analysis of a new gene with WD domains in fish oogenesis".SCIENCE IN CHINA SERIES C-LIFE SCIENCES 44.5(2001):541-553.
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