CAJ | 학술논문

　　以双元载体pPZP100为骨架，通过酶切、连接将标记基因(潮霉素-绿色荧光蛋白，HygR-GFP)及碳色长蠕孢(Helminthosporium carbonum)非核糖体肽合成酶基因(HcNPS6)侧翼序列插入目标载体pPZP100，构建HcNPS6基因敲除载体，并采用冻融法将pPZP100HygR-GFPHcNPS6转入农杆菌菌株AGL-1。挑取经氨苄/氯霉素筛选的重组子进行菌落PCR鉴定。结果表明，敲除载体已成功转入农杆菌AGL-1。
　　이쌍원재체pPZP100위골가，통과매절、련접장표기기인(조매소-록색형광단백，HygR-GFP)급탄색장연포(Helminthosporium carbonum)비핵당체태합성매기인(HcNPS6)측익서렬삽입목표재체pPZP100，구건HcNPS6기인고제재체，병채용동융법장pPZP100HygR-GFPHcNPS6전입농간균균주AGL-1。도취경안변/록매소사선적중조자진행균락PCR감정。결과표명，고제재체이성공전입농간균AGL-1。
Vector construction was based on the skeleton of pPZP100, HygR-GFP and flanking sequences of HcNPS6 of Helminthosporium carbonum were linked into pPZP100 by restriction enzymes digested and ligase connection. Deletion vector of nonribosomal peptide synthetases 6(NPS6) of Helminthosporium carbonum had been constructed pPZP100HygR-GFPHcNPS6 and transformed into AGL-1 by method of freeze-thaw.