cDNA cloning and characterization of a novel SNX gene differentially expressed in previtellogenic oocytes of gibel carp.
To understand the molecular events governing fish oogenesis, a multiple technique was used to identify the genes differentially expressed at different phases during fish oogenesis. This technique is a combination of suppression subtractive hybridization, SMART cDNA synthesis and RACE-PCR. Here we report the cDNA cloning and expression characterization of a novel SNX gene based on its differential transcription between previtellogenic and fully mature oocytes in naturally gynogenetic gibel carp. First, a cDNA fragment selectively expressed in previtellogenic oocytes was identified and used to screen a SMART cDNA library prepared from the same mRNA sample by RACE-PCR for cloning fully length cDNA. The full length cDNA was 1392-bp long and coded for a novel SNX protein with 225 amino acids. The 5' UTR had 72 bp and 3' UTR had 642 bp. Unlike most of maternal genes that are transcribed after vitellogenesis and stored in oocytes, this gene is expressed at a higher level in the previtellogenic oocytes and at a much lower level in fully matured oocytes. However, RT-PCR analysis of tissues showed it was ubiquitous transcription. The novel gene is named fish sorting nexin (fSNX), because it contains a conserved PX domain. The fact which major expression of the gene occurs in the previtellogenic oocytes suggests that it might have an important function in the oogenesis.
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, PR China.
Jianjun
J
Wen
Jing
J
Xie
Jianfan
J
Gui
D000595:Amino Acid Sequence; D000818:Animals; D002347:Carps / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry*; D005260:Female; D029941:Fish Proteins / Q000378:metabolism*; D015870:Gene Expression; D015723:Gene Library; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism*; D016415:Sequence Alignment; D014818:Vitellogenesis / Q000502:physiology*
D018076:DNA, Complementary; D029941:Fish Proteins