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产番茄红素红酵母基因组重排实验条件的优化[J]. 食品工业科技, 2013, (08): 228-231. DOI: 10.13386/j.issn1002-0306.2013.08.032
引用本文: 产番茄红素红酵母基因组重排实验条件的优化[J]. 食品工业科技, 2013, (08): 228-231. DOI: 10.13386/j.issn1002-0306.2013.08.032
Optimizing process of genome shuffling in Rhodotorula sp. producing lycopene[J]. Science and Technology of Food Industry, 2013, (08): 228-231. DOI: 10.13386/j.issn1002-0306.2013.08.032
Citation: Optimizing process of genome shuffling in Rhodotorula sp. producing lycopene[J]. Science and Technology of Food Industry, 2013, (08): 228-231. DOI: 10.13386/j.issn1002-0306.2013.08.032

产番茄红素红酵母基因组重排实验条件的优化

Optimizing process of genome shuffling in Rhodotorula sp. producing lycopene

  • 摘要: 目的:探索基因组重排技术应用于产番茄红素红酵母育种中关键步骤的最优条件。方法:本实验以3株产生番茄红素的红酵母cp-2-11、cp-2-6、cp-28为实验菌株,研究菌体培养时间、酶解浓度、酶解时间、灭活条件以及融合时间对基因组重排的影响。结果:当菌体培养12h,酶解浓度为2%,酶解70min时,原生质体制备与再生率最佳;15W紫外灯,照射距离为35cm,磁力搅拌50r/min的条件下照射18min,60℃下灭活35min作为双亲灭活条件;35%聚乙二醇4000(PEG-4000)作为融合剂,处理23min为最佳融合时间。结论:通过基因组重排最佳条件的确定,后续重排实验的效率得到很大的提升,最终筛选出了高产的目的菌株,产量较融合亲本提高了122%。 

     

    Abstract: Objective:Exploring the optimum conditions of the genome shuffling which applied to the key steps of Rhodotorula. Method:With Rhodotorula cp-2-11、cp-2-6、cp-28 as the tested material, we explored the effects of several factors including cell age, enzyme concentration, hydrolysis time, inactivation conditions, fusion time on genome shuffling. Results:The protoplast preparation rate and regeneration rate was supreme when the cell age was 12 hours, the enzyme concentration was 2% and the hydrolysis time was 70 minute.15W UV light, the irradiation distance was 35cm irradiates 18 minute and 60℃ water bath inactive 35 minutes were used as the conditions of Inactivation, fused 23 minute in 35% PEG as the fusion condition. Conclusions: Through determining the optimum conditions of genome shuffling, the follow-up work was improved effectively. After several rounds of genome shuffling, the high yield strains were breed, which increased by 122% than original strains.

     

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