An improved triploid crucian carp(ITCC) was produced by crossing improved tetraploids(G1×AT,) with improved red crucian carp(IRCC,),which were obtained by distant crossing and gynogenesis.The biological character...An improved triploid crucian carp(ITCC) was produced by crossing improved tetraploids(G1×AT,) with improved red crucian carp(IRCC,),which were obtained by distant crossing and gynogenesis.The biological characteristics of ITCC,including the number and karyotype of chromosomes,gonadad and pituitary structures,phenotype,and growth rate are reported.ITCC possessed 150 chromosomes with the karyotype 33m+51sm+33st+33t.In the breeding season,both ovary-like and testis-like gonads of ITCC were unable to produce normal mature gametes.The ultrastructure of the pituitary of ITCC showed that most of the endocrine granules in gonadotrophic hormone(GTH) cells had not been released,providing endocrinological evidence for the sterility of ITCC.Compared with triploid crucian carp(TCC) produced by mating Japanese crucian carp with allotetraploid hybrids,ITCC not only retained the excellent traits of fast growth rate and sterility,but also acquired improved morphological characteristics,including higher body,shorter tail and smaller head.展开更多
A systemic study was initiated to identify stage-specific expression genes in fish embryogenesis by using suppression subtractive hybridization (SSH) technique. In this study, we presented a preliminary result on scre...A systemic study was initiated to identify stage-specific expression genes in fish embryogenesis by using suppression subtractive hybridization (SSH) technique. In this study, we presented a preliminary result on screen for stage-specific expression genes between tail bud stage (TBS) and heartbeat beginning stage (HBS) in gynogenetic silver crucian carp (Carassius auratus gibelio). Two SSH plasmid libraries specific for TBS embryos and HBS embryos were constructed, and stage-specific expression genes were screened between the two stages. 1963 TBS positive clones and 2466 HBS positive clones were sampled to PCR amplification, and 1373 TBS and 1809 HBS PCR positive clones were selected to carry out dot blots. 169 TBS dot blot positive clones and 272 HBS dot blot positive clones were sequenced. Searching GenBank by using these nucleotide sequences indicated that most of the TBS dot blot positive clones could not be found homologous sequences in the database, while known genes were mainly detected from HBS dot blot positive clones. Of the 79 known genes, 20 were enzymes or kinases involved in important metabolism of embryonic development. Moreover, specific expressions of partial genes were further confirmed by virtual northern blots. This study is the first step for making a large attempt to study temporal and spatial control of gene expression in the gynogenetic fish embryogenesis.展开更多
A 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 natura...A 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 AN-NATG. 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.展开更多
基金Supported by the National Natural Science Fund for Distinguished Young Scholars (Grant No. 30725028)National key Basic Research and Development Program of China (Grant No. 2007CB109206)+2 种基金National Natural Science Foundation of China (Grant No. 30871915)Lotus Scholars Program of Hunan Province,Industrialization Fund of Education Department of Hunan Province (Grant No. 200654-2)Hunan Provincial Innovation Fund for Postgraduates
文摘An improved triploid crucian carp(ITCC) was produced by crossing improved tetraploids(G1×AT,) with improved red crucian carp(IRCC,),which were obtained by distant crossing and gynogenesis.The biological characteristics of ITCC,including the number and karyotype of chromosomes,gonadad and pituitary structures,phenotype,and growth rate are reported.ITCC possessed 150 chromosomes with the karyotype 33m+51sm+33st+33t.In the breeding season,both ovary-like and testis-like gonads of ITCC were unable to produce normal mature gametes.The ultrastructure of the pituitary of ITCC showed that most of the endocrine granules in gonadotrophic hormone(GTH) cells had not been released,providing endocrinological evidence for the sterility of ITCC.Compared with triploid crucian carp(TCC) produced by mating Japanese crucian carp with allotetraploid hybrids,ITCC not only retained the excellent traits of fast growth rate and sterility,but also acquired improved morphological characteristics,including higher body,shorter tail and smaller head.
基金This work was supported by the National Nat-ural Science Foundation of China(Grant No.30130240),the Chinese Academy of Sciences(GrantNo.KSCX2-SW-303).
文摘A systemic study was initiated to identify stage-specific expression genes in fish embryogenesis by using suppression subtractive hybridization (SSH) technique. In this study, we presented a preliminary result on screen for stage-specific expression genes between tail bud stage (TBS) and heartbeat beginning stage (HBS) in gynogenetic silver crucian carp (Carassius auratus gibelio). Two SSH plasmid libraries specific for TBS embryos and HBS embryos were constructed, and stage-specific expression genes were screened between the two stages. 1963 TBS positive clones and 2466 HBS positive clones were sampled to PCR amplification, and 1373 TBS and 1809 HBS PCR positive clones were selected to carry out dot blots. 169 TBS dot blot positive clones and 272 HBS dot blot positive clones were sequenced. Searching GenBank by using these nucleotide sequences indicated that most of the TBS dot blot positive clones could not be found homologous sequences in the database, while known genes were mainly detected from HBS dot blot positive clones. Of the 79 known genes, 20 were enzymes or kinases involved in important metabolism of embryonic development. Moreover, specific expressions of partial genes were further confirmed by virtual northern blots. This study is the first step for making a large attempt to study temporal and spatial control of gene expression in the gynogenetic fish embryogenesis.
基金the Major State Basic Research Program (Grant No. G1999053908) ofChina, the National Natural Science Foundation of China (Grant No.30070402), and the State Key Laboratory of Freshwater Ecology and Biotechnology (Grant No. 2000FB06).
文摘A 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 AN-NATG. 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.
