Construction Of Jatropnha Curcas L. Endosperm CDNA Library And Cloning Research Of Phosphoenolpyruvate Carboxylase Gene

Equal weight endosperm of Jatropha curcas (Linnaeus) at different development stage was used as experimental materials. Total RNA is extracted and purified. Jalropha curcas L. endosperm cDNA library was constructed using cDNA Synthesis Kit, Gigapack III Gold Packaging Extract and Uni-ZAP XR vector. A primary library of 1.5×10^<sup>6Pfu/ml was obtained. An amplified library of 1.5×10¹⁴Pfu/ml was obtained. The recombinant efficiency was 99.7%. The inserted fractions sizes ranged from 0.4kb to 4kb according to the PCR results and sequenced analysis, with an average value of 1kb. After sequenced 10 randomly selected cDNA, Homologous and full-length analysis with other genes indicated that all of picked clones were integrated by using bio-informational methods. The percentage of full-length genes was 20%. This suggests that this library might be suitable to be used to screen full-length gene. These sequences mainly involved in the process of storage protein metabolism, gene expression and their regulation, fatty acid metabolism related genes and signal transduction related genes, including metallothionein-like protein, translation elongation factor and MADS-box gene analog et al. The percentage of storage protein related genes was 20%. The percentage of transcriptional factor related genes was 6%. The percentage of fatty...