产黄青霉突变体库的构建及鉴定-植物学专业毕业论文.docx

河北科技大学硕士学位论文
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万方数据
Abstract
Abstract
Penicillin is one of the important strategic chrysogenum, penicillin producing strain,is an important industrial anism which has great industrial industrial strains are derived from a single natural isolate of P chrysogenum NRRL 1 95 pennicillin production increased by more than 1 000 times.
So it is difficult to improve pennicillin production by traditional breeding and fermentaion
is necessary to obtain a pennicillin hi曲一producing strain by modem molecular biology study is intended to improve the industrial P chrysogenum strain by ic engineering technology to increase the yield of penicillin.
The wildtype strain in our research is industrial P exogenous
bleomycin resistant gene,linked谢th T-DNA which Was synthesized by PCR,was transformed into P chrysogenum protoplast mediated by large number of
transformants were screened by mutants obtained in this way laid the
foundation for screening penicillin high-producing mutant identify if the exogenous gene Was transfomed into P chrysogenum,a simple and high throughput genomic DNA extraction method was method is based on enzyme lysis
combining、Ⅳitll heating and ultrasonic genomic DNA isolated by the
method Can be used as PCR enzymes treatment,6 mg/mL snailase and 4000 U/mL cellulase,is the best way to lyze the mycelia is treated by the following steps,ultrasonic treatment 10—15 min,lyses at 28。C for 14 h,heating at 90-98。C for 10-15 min,then add 50—100 ,mix until the layers separated,The supematant could be used as the PCR template directlyand the PCR bands are clear and accurate.
The mutants were cultivated with 24一well plates screen the pennicillin production
based the high-throughput detection results showed that 324 transforments were identified as mutant strains,includi