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。
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 1 508 bp, including 1 459 bp of an open reading frame with the initiation codon ATG at 45 bp and the termination codon TAA at 1 501 bp and totally encoded 424 amino 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 ligation 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.
出处
《微生物学杂志》
CAS
CSCD
2008年第2期21-26,共6页
Journal of Microbiology
基金
国家高技术研究发展计划(863项目
2006AA10A211)
