作者：胡桂兵, 张上隆, 徐昌杰, 林顺权.
摘要：Based on the codon usage for the amino acids of 177 citrus functional proteins deposited in GenBank, a new antibacterial peptide D (APD) gene with citrus preference codons was designed and chemcially synthesized. The peptide has anability to kill the phloem conducted pathogens for Huanglonbing, a destructive and widespread disease for citrus. The new gene encoding the same peptide, referred to as CAPD, was cloned into pUC19 to generate a recombinant plasmid pUC19-CAPD, and was verified by sequencing, Both pUC19-CAPD and plant expression vector pBI121 were digested by BamH I and Sac I, and then the samller fragment containing CAPD gene from the former and than larger vector fragment without GUS gene from the latter were recovered,ligated, transfromed and identified. As a result, a new plant expression vector, referred to as pHZ05, harboring CAPD traget gene driven by the constitutive 35S promoter of CaMV (35S) was constructed. BamH I and Hind Ⅲ were applied to digest pHZ05 and pUCm-PSP, a plasmid containing phloem specific promoter from Cucubita maxima(PSP), and then the larger vector fragment without 35SP from the former and the smaller fragment containing PSP promoter from the latter were recovered. After ligation, transformation and verification, another new plant expression vector, refered to as pHZ06, harboring CAPD gene driven by different promoters, were then successfully introduced into competent cells of Agrobacterium tumefaciens strains LBA4404, GV3101, EHA105 and A. rhizogenes strains Ri15834, respectively. The study is helpful for further work to find a new way to develop new germplasms resistant to disease Huanglongbing.
关键词：Citrus; Codon optimization; Antibacterial peptide D(AP-D) gene; Phloem specific
Key words: 柑橘;密码子优化;抗菌肽D基因;韧皮部特异启动子;植物表达载体
文献注录：胡桂兵, 张上隆, 徐昌杰, 林顺权.. 韧皮部特异启动子驱动密码子优化的抗菌肽D基因的植物表达载体构建 [J]. 果树学报. 2005, 06: 639-643.
报/刊名：《果树学报》，发表于2005 / 第 06 期。
文献类型： [J]
页码：第 639-643 页 / 共5 页