Wei sheng wu xue bao = Acta microbiologica Sinica · 2009
[Effects of Pa-pex 11 gene on penicillin production in Penicillium aurantiogriseum].
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Yang Jing, An Yang, Xu Xinxin, Liu Gang
Аннотация
OBJECTIVE: To study the effect of Pa-pex 11 gene on penicillin production in Pencillium aurantiogriseum.
METHOD: Based on the conserved domain of pex 11 from Penicillium chrysogenum, we designed primers pex01 and pex02 to amplify Pa-pex 11 gene from P. aurantiogriseum. The complete Pa-pex 11 gene was cloned via thermal asymmetric interlaced PCR (Tail-PCR). In order to obtain tranformants with additional Pa-pex 11 gene copy, ATMT (Agrobacterium tumefaciens mediated transformation) was used to transform P. aurantiogriseum. The copy number of Pa-pex 11 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. aurantiogriseum. Increasing the copy number of Pa-pex 11 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-pex 11 can increase the number of microbodies, and stimulate the penicillin production.
METHOD: Based on the conserved domain of pex 11 from Penicillium chrysogenum, we designed primers pex01 and pex02 to amplify Pa-pex 11 gene from P. aurantiogriseum. The complete Pa-pex 11 gene was cloned via thermal asymmetric interlaced PCR (Tail-PCR). In order to obtain tranformants with additional Pa-pex 11 gene copy, ATMT (Agrobacterium tumefaciens mediated transformation) was used to transform P. aurantiogriseum. The copy number of Pa-pex 11 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. aurantiogriseum. Increasing the copy number of Pa-pex 11 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-pex 11 can increase the number of microbodies, and stimulate the penicillin production.