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首页> 外文学位 >ISOLATION AND CHARACTERIZATION OF GENES FROM BETA-LACTAM ANTIBIOTIC PRODUCING ORGANISMS (PENICILLIUM CHRYSOGENUM, RIBOSOMAL-RNA, CEPHALOSPORIUM ACREMONIUM, ISOPENICILLIN N SYNTHETASE).
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ISOLATION AND CHARACTERIZATION OF GENES FROM BETA-LACTAM ANTIBIOTIC PRODUCING ORGANISMS (PENICILLIUM CHRYSOGENUM, RIBOSOMAL-RNA, CEPHALOSPORIUM ACREMONIUM, ISOPENICILLIN N SYNTHETASE).

机译：从β-内酰胺类抗生素生产基因（青霉菌，核糖体RNA，头孢杆菌，异戊二烯N合成酶）中分离和鉴定基因。

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摘要

The potential exists to increase the production of existing beta-lactam antibiotics and to invent new antibiotics by genetic engineering of Penicillium chrysogenum and Cephalosporium acremonium. The purpose of this study was to further characterize the genetics of these fungi and to isolate and characterize genes so that they could be manipulated to increase beta-lactam production and create new antibiotics.; The isopenicillin N synthetase gene from P. chrysogenum and the rRNA genes from C. acremonium were isolated from recombinant lambda bacteriophage libraries using heterologous probes. The protein coding region of the P. chrysogenum isopenicillin N synthetase gene was about 74% homologous to the C. acremonium isopenicillin N synthetase gene, and the predicted amino acid sequences of the encoded proteins were about 73% homologous. E. coli cells with the P. chrysogenum isopenicillin N synthetase gene contained isopenicillin N synthetase activity whereas untransformed cells were completely devoid of this enzymatic activity. The transformed cells were also shown to contain an abundant protein accounting for about 10% of total cell protein which reacted strongly with anti - C. acremonium isopenicillin N synthetase antiserum.; The rRNA genes of C. acremonium were located on a 9.2 kb PstI restriction fragment in genomic DNA digests. A partial restriction map was generated of the 9.2 kb rDNA repeat and hybridization analysis revealed the order of the genes to be 18S, 5.8S and 28S. Ribosomal DNA sequences were incorporated into C. acremonium transformation vectors to increase the transformation frequency and thus facilitate identifying a high producing strain.
机译：存在通过潜在的产黄青霉和顶头孢霉的基因工程来增加现有β-内酰胺抗生素的生产和发明新抗生素的潜力。这项研究的目的是进一步表征这些真菌的遗传，并分离和表征基因，以便可以操纵它们以增加β-内酰胺的产生并创造新的抗生素。使用异源探针从重组λ噬菌体文库中分离出产黄青霉的异青霉素N合成酶基因和顶头孢霉的rRNA基因。产黄青霉异青霉素N合成酶基因的蛋白编码区与顶头孢霉异青霉素N合成酶基因的同源性约为74％，编码蛋白的预测氨基酸序列同源性约为73％。具有产黄青霉异黄霉素N合成酶基因的大肠杆菌细胞含有异青霉素N合成酶活性，而未转化的细胞完全没有这种酶活性。还显示出转化的细胞含有丰富的蛋白质，约占总细胞蛋白质的10％，该蛋白质与抗顶头孢霉异青霉素N合成酶抗血清强烈反应。顶头孢霉的rRNA基因位于基因组DNA消化物中的9.2kb PstI限制性片段上。产生了9.2kb rDNA重复序列的部分限制性图谱，杂交分析揭示了基因的顺序为18S，5.8S和28S。将核糖体DNA序列掺入顶头孢霉转化载体中以增加转化频率并因此有助于鉴定高产菌株。

著录项

  • 作者

    CARR, LUCINDA GAYLE.;

  • 作者单位

    Indiana University.;

  • 授予单位 Indiana University.;
  • 学科 Biology Molecular.
  • 学位 Ph.D.
  • 年度 1986
  • 页码 134 p.
  • 总页数 134
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;
  • 关键词

  • 入库时间 2022-08-17 11:51:04

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