文献类型： 会议论文
第一作者： Zhang Shuzheng
作者： Zhang Shuzheng 1 ; Zheng Xueqin;
作者机构： 1.National Key Biotechnology Laboratory for Tropical Crops, CATAS, Haikou, Hainan, China 571101
关键词： Trehalose synthase gene;Cloning;Sugarcane;Transformation
会议名称： Promoting global innovation of agricultural science & technology and sustainable agriculture development,Session 4
主办单位：
页码： 123-128
摘要： The total RNA from bacidiomycete, Grifola frondosa was isolated. The mRNA was purified and reverse transcripted to first strand of cDNA. A pair of primer was desiqued and synthesized according to the sequence reported. The very specific PCR product oftrehalose synthase gene was obtained by using the first strand of cDNA as template. The product was ligased to pGEM~(R)-T easy vector and sequenced. The sequencing data showed that the PCR product was 2199bp, and contained a start codon and a stop codonIt encoded 732 amino acids and was an intact open reading frame. Comparation of the cDNA sequence from this experiment with that reported by Saito., K.(1998) indicated the homology was 98.6%. A series of trehalose synthase gene expression vectors were constructed by using different promoters. The pBT was was constructed by inserting the trehalose synthase gene between the CaMV 35S promoter and NOS terminator into the expression vector pBI1 21 and pUBT was constructed by using ubi-L promoter and NOS terminator. Then the pBT and pUBT were introduced into sugarcane via microprojectile bombardment. The transformants were selected by geneticin. PCR amplification and Southern analysis showed that the Tsase gene was integrated into sugarcane genomes.The RNAdot blotting analysis revealed that the Tsase gene was expressed in transcriptional level in sugarcane.
分类号： S
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