【摘要】 利用PCR技术从绿色木霉LTR-2(Trichoderma virede)基因组DNA中扩增到一段序列,测序结果表明,该编码基因片段大小为1508 bp,其中包括一个1459 bp的开放阅读框,起始密码子位于45 bp,终止密码子位于1501 bp,共编码氨基酸424个。在Genbank中进行序列比对,发现该序列同已发表的Trichoderma viride 42 ku几丁质酶氨基酸序列具有99%的同源性。将该片段同pCAMBLA1300中的35S启动子和35S-polyA终止子连接后,插入载体pCAMBIA1302多克隆位点中,构建成植物转化载体,最后将构建好的载体pCHI1302- 42通过转化导入根癌农杆菌LBA4404中,为进一步构建转基因植物奠定了基础。更多还原

【Abstract】 A piece of genome DNA from Trichoderma viride LTR-2 was cloned using PCR amplification,the results of sequencing indicated that the size of the encoded gene fragment was 1508 bp,including 1459 bp of an open reading frame with the initiation codon ATG at 45 bp and the termination codon TAA at 1501 bp and totally encoded 424 ami- no acids.It was found that in the sequence comparison with Genbank the sequence was homologous at 99% between amino acid sequence of chitinase of Trichoderma viride 42 ku that has been reported and the sequence.After the liga- tion of the sequence with 35S promoter and 35S-polyA terminator from pCAMBIA1300 then inserted into carrier pCAMBIA1300 poly-cloned site to construct plant transformation carrier,finally the constructed carrier pCHI1302-42 was introduced into Agrobacterium tumefaciens LBA4404 to lay a foundation for further construction of transgenic plant.更多还原