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Aspergillus fumigatus에서 Methyltransferase 유전자 AfuvipB와 AfuvipC의 분리 및 분석

모하메드 ( Mohammed A. Abdo Elgabbar ),한갑훈 ( Kap Hoon Han ) 한국균학회 2015 Mycobiology Vol.43 No.1
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In filamentous fungi, velvet complex associated with the veA gene plays pivotal roles in development and secondary metabolism. In a model fungus Aspergillus nidulans, many proteins that can interact with VeA, including two methyltransferases VipB and VipC, have been isolated and characterized. In this study, we isolated homologs of the vipB and vipC genes in the human opportunistic pathogenic fungus Aspergillus fumigatus and named AfuvipB and AfuvipC. The AfuvipB gene, annotated as Afu3g14920 in the Aspergillus Genome Database (AspGD) database, consists of 1,510 bp interrupted with 10 introns yielding 336 amino acid-long putative methyltransferase protein. Similarly, AfuvipC, which is Afu8g01930, has 10 introns and encodes a polypeptide with 339 amino acids having a methyltransferase domain in the middle of the protein. To characterize the function of the genes in A. fumigatus, knock-out mutants were generated and the phenotypes were observed. Deletion of AfuvipB gene caused no obvious phenotypic change on point inoculation but showed smaller colony than wild-type when the mutant was subjected to culture on single spore-driven culture condition. However, AfuvipC deletion mutant demonstrated no phenotypic difference from wild type both in point inoculation and streaking cultures. These results indicate that the two methyltransfereases might have a redundant role and could be dispensable in normal culture conditions.
2
순창군 장류로부터 분리된 황국균의 동정 및 특성

임은미 ( Eun Mi Lim ),이지영 ( Ji Young Lee ),모하메드 ( Mohammed A. Abdo Elgabbar ),한갑훈 ( Kap Hoon Han ),이보순 ( Bo Soon Lee ),조용식 ( Yong Sik Cho ),김현영 ( Hyoun Young Kim ) 한국균학회 2014 韓國菌學會誌 Vol.42 No.4
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  ScienceON
  
  KISS
In this study, we attempted to isolate fungi from soybean fermented foods produced in Sunchang County and to identify Aspergillus oryzae from fungal isolates. Ten fungal isolates were identified with β-tubulin gene. According to the sequences of β-tubulin gene, ten fungal isolates were identified as A. oryzae/flavus complex. For further identification of the ten of fungal isolates, omtA gene, one gene of the aflatoxin biosynthesis gene cluster, was sequenced and the sequences were compared with those of A. oryzae and A. flavus strains from the GenBank database. In addition, identification of the ten fungal isolates was further confirmed using the PCR amplicon of norB and cypA intergenic region, in which a deletion was recognized relative to A. flavus and A. parasiticus. The amplicon size of the ten fungal isolate strains was smaller than those of A. flavus and A. parasiticus, but the same as that of the reference A. oryzae strain. These results indicated that the ten isolates should be identified as A. oryzae. The protease activity in rice koji made with 6, 13, 17, 27, 37 and 38 of strain, respectively was twice higher than that in control. The kojis made with nine of the A. oryzae isolates, respectively, did not produce aflatoxin, suggesting that the strains could possibly be used as starters for soybean products.
3
Aspergillus fumigatus에서 Methyltransferase 유전자 AfuvipB와 AfuvipC의 분리 및 분석

모하메드,한갑훈,Elgabbar, Mohammed A. Abdo,Han, Kap-Hoon 한국균학회 2015 韓國菌學會誌 Vol.43 No.1
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사상성 진균에서 veA 유전자와 연계되어 있는 velvet 복합체는 진균의 분화와 이차 대사산물의 조절에 매우 중요한 기능을 한다. 모델 사상균인 Aspergillus nidulans의 경우 methyltransferase인 VipB와 VipC를 포함한 여러 단백질들이 VeA 단백질과 상호작용하는 것으로 알려져 있다. 본 연구에서는 인간 기회감염 진균인 Aspergillus fumigatus에서 vipB와 vipC 유전자의 상동유전자를 분리하여 각각 AfuvipB와 AfuvipC로 명명하였다. AfuvipB 유전자는 AspGD 데이터베이스에 Afu3g14920으로 등록되어 있으며 1,510 bp 길이에 10개의 인트론을 가지고 있고, 유전자 산물은 336 아미노산 잔기로 구성된 단백질로 methyltransferase 도메인을 가지고 있었다. AfuvipC는 Afu8g01930으로 AfuvipB와 유사하게 10개의 인트론을 가지고 있으며 339개의 아미노산으로 구성된 methyltransferase를 암호화하고 있었다. A. fumigatus에서 각각의 유전자에 대한 기능을 알아보고자 유전자제거 돌연변이 균주들을 제조하고 그들의 표현형을 관찰하였다. AfuvipB 유전자 제거 돌연변이는 점 접종을 하였을 경우 대조군에 비하여 표현형의 차이를 보이지 않았다. 그러나 단일 포자에서 성장한 콜로니를 비교해 보았을 때 대조군에 비하여 그 크기가 작고 분화 속도도 약간 더딘 것을 관찰할 수 있었다. 반면에 AfuvipC 유전자 제거 돌연변이는 대조군과 비교하였을 때 표현형의 차이를 보이지 않았다. 이러한 결과는 두 개의 methyltransferase가 상호 중복적인 역할을 수행하거나 정상적인 실험실 배양조건에서는 중요한 기능을 수행하지 않을 수 있음을 시사한다. In filamentous fungi, velvet complex associated with the veA gene plays pivotal roles in development and secondary metabolism. In a model fungus Aspergillus nidulans, many proteins that can interact with VeA, including two methyltransferases VipB and VipC, have been isolated and characterized. In this study, we isolated homologs of the vipB and vipC genes in the human opportunistic pathogenic fungus Aspergillus fumigatus and named AfuvipB and AfuvipC. The AfuvipB gene, annotated as Afu3g14920 in the Aspergillus Genome Database (AspGD) database, consists of 1,510 bp interrupted with 10 introns yielding 336 amino acid-long putative methyltransferase protein. Similarly, AfuvipC, which is Afu8g01930, has 10 introns and encodes a polypeptide with 339 amino acids having a methyltransferase domain in the middle of the protein. To characterize the function of the genes in A. fumigatus, knock-out mutants were generated and the phenotypes were observed. Deletion of AfuvipB gene caused no obvious phenotypic change on point inoculation but showed smaller colony than wild-type when the mutant was subjected to culture on single spore-driven culture condition. However, AfuvipC deletion mutant demonstrated no phenotypic difference from wild type both in point inoculation and streaking cultures. These results indicate that the two methyltransfereases might have a redundant role and could be dispensable in normal culture conditions.