Cloning of CCoA OMT gene in populus deltoides and construction of antisense plant expression vector
2008
Xia Qiongmei | Tian Min | Fan Zhengqi
Chinese. 据已报道的CCoA OMT（caffeoyl-CoA3-O-methyltransferase）基因的cDNA序列设计一对兼并引物，从美洲黑杨当年生嫩枝的次生木质部中提取总RNA，通过RT-PCR的方法获得约750bp的特异性扩增产物。测序结果显示，克隆的序列与GenBank中注册的CCoA OMT基因cDNA同源性最高达99％。该基因的cDNA序列已在GenBank上登录（登录号：EF153198）。将克隆的美洲黑杨CCoA OMT基因的cDNA序列反向连接到植物表达载体pBI121中CaMV 35S启动子的下游，成功构建了美洲黑杨CCoA OMT基因的反义表达载体pBI-CCoAOMT，为进一步通过反义技术降低杨树木质素含量奠定了基础。
Show more [+] Less [-]English. A pair of primers was designed according to the reported cDNA sequence of CCoA OMT （Caffeoyl-CoA 3-O-methyltransferase） gene, the total RNA was extracted from the developing second xylem of Populus deltoides, and an amplification product about 750 bp was obtained by RT-PCR reaction. Sequence analysis showed that the cloned cDNA shares a homology 99% in comparison with the reported data. This sequence has been registered in GenBank （Accession number： EF153198）. An antisense expression vector, named pBI-CCoAOMT, of the CCoA OMT gene was constructed, in which the antisense sequence was controlled by the CaMV 35S promoter. The work laid the foundations of the future transgenic research for reducing lignin content of poplar via antisense technology.
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