Screening of growth- or development-related genes by using genomic library with inducible promoter in Aspergillus nidulans.
Using the genomic library constructed at the downstream of the niiA promoter, which induces the over-expression of an inserted DNA fragment, we have attempted to screen the genes affecting growth or development by over-expression. The wild-type strain was transformed using the AMA-niiA(p) library and cultured on 1.2 M sorbitol media, in which asexual sporulation is induced, but sexual development is repressed. Over 100,000 strains transformed to pyrG(+) were analyzed with regard to any changes in phenotype. Consequently, seven strains were isolated for further analyses. These strains were designated NOT [niiA(p) over-expression transformants] stains. Four of the strains were of the inducible type, and the remaining strains were of the multi-copy suppression type. Two of the inducible-type strains, NOT1 and NOT40, harbored genes which had been inserted in reverse direction, suggesting that the mutant phenotypes had been derived from an excess amount of anti-sense mRNA. Domain analyses of the deduced polypeptides from the DNA fragments rescued from the transformants revealed that NOT1, NOT40 and NOT6 harbored a LisH motif, a forkhead domain, and a Zn(II)(2)Cys(6) binuclear zinc cluster, respectively.
Lee
Bang-Yong
BY
Department of Biology, Graduate School of Wonkwang University, Iksan, Republic of Korea.
Han
Sang-Yong
SY
Choi
Han Gil
HG
Kim
Jee Hyun
JH
Han
Kap-Hoon
KH
Han
Dong-Min
DM
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000166:cytology / Q000235:genetics* / Q000254:growth & development; D017124:Conserved Sequence; D018628:Gene Dosage; D015870:Gene Expression; D015723:Gene Library; D005800:Genes, Fungal; D005822:Genetic Vectors; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D009024:Morphogenesis / Q000235:genetics; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary / Q000235:genetics; D016372:RNA, Antisense; D016415:Sequence Alignment
D016372:RNA, Antisense