灰黄青霉Pa-pex11基因对青霉素产生的影响

Effects of Pa-pex11 gene on penicillin production in Penicillium anrantiogriseum

摘要:

摘要：【目的】研究灰黄青霉Pa-pex11对青霉素产生的影响。【方法】通过保守序列设计引物从灰黄青霉中克隆了含有pex11同源基因DNA片段，进而通过热不对称交错PCR(Thermal asymmetric interlaced PCR, Tail PCR )扩增得到灰黄青霉中Pa-pex11基因的全序列。利用根癌农杆菌介导的遗传转化（ATMT）系统，在灰黄青霉基因组上导入额外的Pa-pex11基因。对灰黄青霉野生型菌株和转化子进行了青霉素发酵及生物活性测定。通过荧光定量PCR(quantitative

Abstract:

Abstract: [Objective] To study the effect of Pa-pex11 gene on penicillin production in Pencillium anrantiogriseum. [Method] Based on the conserved domain of pex11 from Penicillium chrysogenum, we designed primers pex01 and pex02 to amplify Pa-pex11 gene from P. anrantiogriseum. The complete Pa-pex11 gene was cloned via thermal asymmetric interlaced PCR (Tail-PCR). In order to obtain tranformants with additional Pa-pex11 gene copy, ATMT (Agrobacterium tumefaciens mediated transformation) was used to transform P. anrantiogriseum. The copy number of Pa-pex11 was measured by quantitative PCR. Penicillin production of the transformant was analyzed by bioassays and the microbodies were observed by transmission electron microscope (TEM). [Results] ATMT was successfully applied in P.anrantiogriseum. Increasing the copy number of Pa-pex11 gene resulted in a 1.7-fold increase of pencillin production, and the electron microscope graph showed that the number of microbodies increased obviously. [Conclusion] Our results suggest that increasing the copy number of Pa-pex11 can increase the number of microbodies, and stimulate the penicillin production.