Aspergillus niger로 부터 $\alpha$-glucosidase 발현억제 형질전환체의 분리

  • 이동건 (한국과학기술연구원 생명공학연구소) ;
  • 이진영 (한국과학기술연구원 생명공학연구소) ;
  • 서영배 (경북대학교 미생물학과)
  • Published : 1996.08.01

Abstract

We have already cloned an extracellular $\alpha$-glucosidase gene from Aspergillus niger with oligonucleotide probe synthesized on the basis of the peptide sequences determined previously. The DNA sequence revealed an open reading frame of 895 amino acids split by three introns. We are attempting to construct an A. niger strain deficient in the $\alpha$-glucosidase enzyme activity, which would be useful for the glucoamylase production without contamination by the industrially undesirable $\alpha$-glucosidase. For destruction of the $\alpha$-glucosidase gene, we try to make transformations. A cloned partial $\alpha$-glucosidase gene was introduced into Aspergillus niger, and transformants with suppressed $\alpha$-glucosidase activity were isolated. The transformants were cultured on YPD medium which contained Hygromycin B at 30$\circ$C. The activity of $\alpha$-glucosidase of the suppressed transformants was compared to that of wild type activity. As shown by southern-hybridization, we detected that the transformant was a heterocaryon.

Keywords

References

  1. Enzyme Hanbook v.Ⅱ Barman.T.E.
  2. J. Brew. Soc. Japan. v.72 Enzyme in sake making IX. Properties of fungal transglucosidase Iwano.K.;K.Shibata.;K.Mitsunage.;Y.Nunokawa.
  3. J. Brew. Soc. Japan. v.72 Enzyme in sake making. VII. Estimation of transglucosidase in the presence of α-amy-lase and glucoamylase Iwano.K.;M.Sato;K.Shibata.;K.Mitsunaga.;Y.Nunokawa
  4. Gene v.57 Transformation of Aspergillus nidulans with the hygromycin resistance gene Cullen.D.;S.A.Leong.;L.T.Wilson.;D.J.Henner
  5. Agric. Biol. Chem. v.51 Integrative transformation of Aspergillus oryzae with a plasmid containing the Aspergillus nidulans argB gene Gomi.K.;Y.Iimura.;S.Hara
  6. J. Biotechnol. Cloning and sequencing of an α-glucosidase gene from Aspergillus niger and its expression in A. nudulans Nakamura.A.;I.Nishimura.;A.Yokoyama.;D.G.Lee.;M.Hidaka.;H.Masaki.;A.Kimura.;S.Chiba.;T.Uozumi