利用重叠PCR技术合成杂合抗菌肽基因CB-Tmp1

Synthetic heterozygous antibacterial peptides CB-Tmp1 by overlapping PCR technology

以Cecropin B与Tmp1为母本,设计了两条杂合抗菌肽。由于直接合成基因全序列费用比较高,为降低实验成本,实验采用重叠PCR法。将两条杂合抗菌肽基因序列总共分成7个部分,设计了6条重叠引物,利用重叠PCR法成功合成抗菌肽基因全序列,并构建了重组质粒pMD18-T1和pMD18-T2。经过测序验证,利用重叠PCR的方法扩增出来的基因序列与设计的序列完全相符。

The experiments designed two heterozygous antibacterial peptides which composed by Cecropin B and Tmp 1. Because of the direct synthesis genes were higher than the cost of using the method of overlapping PCR,this experiments used the method of overlapping PCR to synthesis heterozygous antibacterial peptides. The two heterozygous genes were separated into seven parts and designed 6 overlap primers. Used overlapping PCR antibacterial peptides synthesised the gene sequences,and then constructed th~ ecombinant plasmid pMD18-T1 and pMD18-T2. Finally,sequencing identified the sequences were correct.