为探讨HIV-Tat蛋白转导域(protein transducfion domain,PTD)对中国鲎抗菌肽tachyplesin的协同作用,试验以经毕赤酵母密码子优化后的tachyplesin成熟肽(54aa)加TAT PTD(11 aa)的cDNA序列为参考模板,设计了6条单链寡核苷酸片段,首尾引物的5'端分别引入EcoR Ⅰ和XMⅠ酶切位点,通过重叠延伸PCR方法成功合成了TAT PTD+ Tachyplesin序列,序列全长219 bp,为其后续功能与协同作用研究奠定了基础.%In order to study the coefficient of the TAT PTD (the protein transduction domain in HIV-1 transactivator of transcription protein) to the antibacterial peptide tachyplesin from Tachpleus tridentatus, 6 sequence of oligonucleotides with EcoR land Xba I in the ends of 5'-end of the first and last primers respectively were designed, based on the TAT PTD and tachyplesin mature peptide cDNA template, which was optimized according to the Pichia pastoris preferred codon. A optimized TAT PTD and tachyplesin fusion gene with 219 base pairs was artificially synthesized by overlap extension PCR, which laid a preliminary foundation for the following functional and coefficient studies.
展开▼
