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当前位置: 首页 > 文章 > 以吡啶硫胺素抗性基因为选择标记的红曲菌原生质体转化研究 安徽农业科学 2012,40 (23) 65-67
Position: Home > Articles > Transformation of Monascus aurantiacus AS3.4384 with Pyrithiamine Resistance Gene(ptrA)as a Dominant Selectable Marker Journal of Anhui Agricultural Sciences 2012,40 (23) 65-67

以吡啶硫胺素抗性基因为选择标记的红曲菌原生质体转化研究

作  者：
崔华;李燕萍
单  位：
南昌大学食品科学与技术国家重点实验室;南昌大学中德联合研究院
关键词：
红曲菌;吡啶硫胺素抗性;原生质体;转化
摘  要：
[目的]以吡啶硫胺素抗性基因(Pyrithiamine Resistance Gene,ptrA)为选择标记基因,对红曲菌原生质体转化体系进行研究,以期为红曲菌基因功能的研究奠定基础。[方法]采用PEG-CaCl2介导的原生质体转化法,将含有吡啶硫胺素抗性基因(ptrA)的pPTRⅡ质粒转入橙红色红曲菌AS3.4384原生质体中,随机挑选5个转化子,通过PCR方法和Southern杂交对转化结果进行检测,验证pPTRII质粒是否整合到转化子染色体DNA中。[结果]pPTRⅡ质粒转入红曲菌AS3.4384原生质体中的转化率为20～24个/μg,PCR检测结果显示,基因组DNA中有吡啶硫胺素抗性基因的存在,Southern杂交分析发现,ptrA基因随机整合到了转化子的染色体上,该结果表明ptrA基因可用作红曲菌转化的选择标记。[结论]该研究是吡啶硫胺素抗性基因在红曲菌中的首次转化研究,试验结果为红曲菌的基因功能研究奠定了基础。
译  名：
Transformation of Monascus aurantiacus AS3.4384 with Pyrithiamine Resistance Gene(ptrA)as a Dominant Selectable Marker
作  者：
CUI Hua et al(State Key Laboratory of Food Science and Technology,Sino-Germany Joint Research Institute,Nanchang University,Nanchang,Jiangxi 330047)
关键词：
Monascus aurantiacus;Pyrithiamine resistance;Protoplast;Transformation
摘  要：
[Objective] To study the transformation of protoplasts of the strain Monascus aurantiacus AS3.4384 by using pyrithiamine resistance gene(ptrA)as the selectable marker,so as to lay foundation for studying the gene functions of Monascus.[Method] Plasmid of pPTRⅡwas transformed by PEG-CaCl2 method.The pPTR II plasmid containing ptrA was transfected into the AS3.4384 protoplast,five transformants were randomly selected and the transformation results were detected by PCR method and Southern blot analysis,it was proved that the plasmid of pPTRⅡ had been integrated into the chromosome DNA.[Result] The transformation rate of pPTR II in AS3.4384 was 20-24 transformants/μg,the PCR detection results indicated that ptrA existed in DNA,Southern blot analysis indicated that the ptrA gene was integrated into the chromosome of transformants,the above results proved that the ptrA gene could be a selective marker in the transformation of M.aurantiacus.[Conclusion] This is the first report for the transformation of ptrA in Monascus and lays a foundation for studying the gene function of M.aurantiacus.

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