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首页> 外文会议>国际营养科学联盟第八届临床营养学大会暨第五届亚太临床营养学会大会 >Construction of a replacement vector to disrupt pksCT gene for the mycotoxin citrinin biosynthesis in Monascus aurantiacus and maintain food red pigment production
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Construction of a replacement vector to disrupt pksCT gene for the mycotoxin citrinin biosynthesis in Monascus aurantiacus and maintain food red pigment production

机译：构建替代载体以破坏红曲霉菌中真菌毒素柠檬素生物合成的pksCT基因并维持食物红色素的产生

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More and more people pay attention to citrinin produced by Monascus, which has nephrotoxic activity in mammals. It was reported that pksCT gene is responsible for citrinin biosynthesis in Monascus purpureus. In this paper, two DNA fragments in both ends of pksCT were amplified by genomic PCR from fourteen Monascus spp.strains. The PCR products were gained from all of the strains. It is suggested that pksCT gene was highly conserved in different citrinin-producing Monascus strains. A pksCT-replacement vector (pHD 106) was constructed to disrupt pksCTwith a hygromycin resistance gene as the selection marker, and was transformed into M. aurantlacus Li AS3.4384. Three transformants (M. aurantiacus PHDS 18, PHDS26, PHDS31) were selected from transformant selective plates. The targeting fragment D was gained by genomic PCR from PHDS18 and PHDS26 except PHDS31. The expressing citrinin capacities of PHDS26 was decreased by about 98%, while PHDS18 was reserved the high capacity of producing citrinin, after 10 days of growth on YM medium. The results indicated that PHDS26 is a pksCT-disrupted strain. There are maybe other genes besides pksCTresponsible for citrinin biosynthesis in M. aurantiacus. It is the effective way to solve the problem of citrinin in M. aurantiacusproducts by constructing replacement vectors to disrupt the genes responsible for citrinin biosynthesis to reduce the capacity of expressing citrinin.
机译：越来越多的人关注由红曲霉产生的柑桔素，其在哺乳动物中具有肾毒性。据报道，pksCT基因负责紫癜红霉素中的柠檬素生物合成。本文通过基因组PCR从14个红曲菌菌株中扩增了pksCT两端的两个DNA片段。从所有菌株获得PCR产物。提示pksCT基因在不同的生产柠檬绿素的红曲菌菌株中高度保守。构建了以潮霉素抗性基因为选择标记的pksCT置换载体（pHD 106），以破坏pksCT，并将其转化为金黄色葡萄球菌AS3.4384。从转化体选择板中选择了三个转化体（M. aurantiacus PHDS 18，PHDS26，PHDS31）。通过基因组PCR从PHDS31和PHDS31获得靶向片段D。在YM培养基上生长10天后，PHDS26的表达citrinin能力下降了约98％，而PHDS18保留了高产量的生产citrinin的能力。结果表明，PHDS26是pksCT破坏的菌株。除了pksCT以外，可能还有其他基因可用于奥兰蒂斯酵母中的柠檬素生物合成。通过构建替代载体来破坏负责柑桔素生物合成的基因以降低表达柑桔素的能力，是解决金黄色葡萄球菌产品中的柑桔素问题的有效途径。

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