Jatropha (jatropha Curcas L.) Endosperm Cdna Library Screening Analysis And Jc Of The-fbl1 Genes, Jc Of The-tctp1 Gene, Cloning Research

50 randomly selected cDNA clones from Jatropha curcas(Linnaeus) endosperm cDNA library were sequenced and 47 expressed sequence tags(ESTs) information were obtained. With Blastn and Blastx analysis, these ESTs were further grouped into 10 functional categories including: metabolism(10.64％), cell defence(8.51％), protein synthesis(17.02％), energy(638％), transport facilitation(4.26％), signal transduction(8.51％), cell growth, division(2.13％), transcription(4.26％), cell structure(4.26％) and function unknown(34.04％).Among the 47 ESTs, a new gene was cloned and named as Jc-Fbll. The cloned cDNA encompasses a 1224bp open reading frame, which encodes a 407 amino acid peptide(GenBank ID: DQ900595). Alignments with F-box proteins in plants show 52％and 75％identity with Arabidopsis thaliana and Oryza sativa(japonica cultivar-group) respectively; while alignments with F-box proteins in animals show 48％～56％identity. Pfam search predicted some conserved domains or motifs in the deduced protein, including F-box and leucine-rich repeat domains. Genomic gene sequence analysis indicated two extrons and an intron with the length of 331bp, 893bp, and 362bp respectively. The similar genomic character was also found in the Arabidopsis thaliana and Oryza sativa homologues. Moreover, an eukaryotic expression vector, pBI121-Jc-Fbll-GFP, was constructed and confirmed for farther research work.Also from this cDNA library, another gene was cloned and named as Jc-TCTP1. Jc-TCTP1 cDNA encompasses an open reading frame with 507bp, which encodes a 168 amino acid peptide(GenBank ID: EF091818). Alignments with translational controlled tumor proteins(TCTP) in plants show 93％, 90％, 84％and 79％identity with Hevea brasiliensis, Elaeis guineensis, Oryza sativa(japonica cultivar-group) and Arabidopsis thaliana respectively. Pfam search and Prosite search predicted some conserved domains or motifs in the deduced protein, including a TCTP domain. Genomic gene sequence analysis indicated five extrons and four introns. Besides, two eukaryotic expression vectors, pBI121-Jc-TCTP1-1 and pBI121-Jc-TCTP1-2, were constructed and confirmed, the former one only contains Jc-TCTP1 ORF, while the later one contains Jc-TCTP1 ORF along with its 5'-UTR.