Cloning of cDNA 5’End of Gonad Differential Expression Gene GD13 in Penaeus japonicus
Jiaoping Zhang
Abstract
Jiaoping Zhang
Abstract
In this paper,the method of SMART Rapid Amplification of cDNA ends (RACE) was employed to clone the 5’ end of cDNA of GD13,which is a gene differentially expressed between ovary and testis of Penaeus japonicus.The SMARTII Oligo nucleotide was used as a template to stretch the cDNA 5’end when the first strand cDNA was synthesized.Then a Nested-PCR was carried out to specially amplify the interested cDNA with the adaptor primers (UPM and NUP) and the gene special primers (13GSP1 and 13GSP2).As a result,we got a 656 bp full length cDNA of GD13 which encodes 164 amino acid.Homologous comparison of amino acid sequence demonstrated that this gene shared significant homology with the ribosome protein L24 gene of Branchiostoma belcheri,Mus musculus and L30 gene of Bos Taurus.Further work investigating why this ribosome protein gene differentially expressed in testis and ovary of shrimp is going on.
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In this paper,the method of SMART Rapid Amplification of cDNA ends (RACE) was employed to clone the 5’ end of cDNA of GD13,which is a gene differentially expressed between ovary and testis of Penaeus japonicus.The SMARTII Oligo nucleotide was used as a template to stretch the cDNA 5’end when the first strand cDNA was synthesized.Then a Nested-PCR was carried out to specially amplify the interested cDNA with the adaptor primers (UPM and NUP) and the gene special primers (13GSP1 and 13GSP2).As a result,we got a 656 bp full length cDNA of GD13 which encodes 164 amino acid.Homologous comparison of amino acid sequence demonstrated that this gene shared significant homology with the ribosome protein L24 gene of Branchiostoma belcheri,Mus musculus and L30 gene of Bos Taurus.Further work investigating why this ribosome protein gene differentially expressed in testis and ovary of shrimp is going on.
Key concepts: Complementary DNA, Biology, Molecular biology, Gene, Rapid amplification of cDNA ends, Homology (biology), cDNA library, Gene expression