• Applied Biological Chemistry
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• 제11권
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• pp.137-141
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• 1969

Amino 산(酸) 핵산관련물질등(核酸關連物質等)을 발효생산(醱酵生産)하는데 응용(應用)키 위(爲)하여 Adenine auxotrophic mutants를 분리(分離)하려고 Gram positive이며 Spore forming bacteria 인 Bacillus subtilis 및 Gram negative bacteria 인 Escherichia coli 를 시험균주(試驗菌株)로 하였다. 이들 균주(菌株)에 U.V-light 처리(處理)와 그리고 Vitamin 및 핵산(核酸) Analogue와 Aminopterine, Streptomycin 처리(處理)로서 효율(效率)좋게 Adenine auxotrophic mutants를 분리(分離)하였다. 1. U.V. -light$(2530\;{\AA}\;2080\;erg/mm^2)$, distance 30 cm) $80{\sim}90sec$. 조사(照射)가 Bacillus subtilis의 Auxotrophic mutants 4주(株) 그리고 E. coli에 $15{\sim}20sec$. 조사(照射)는 Auxotrophic mutants 8주(株)를 분리(分離)하는데 최적조건(最適條件)임을 알았다. 2. Bacillus subtilis 에 대(對)하여 위의 최적조건(最適條件)으로 조사(照射)하는데 Aminopterine$(200\;{\mu}g/ml)$. 처리구(處理區)가 생육(生育)을 현저(顯著)히 조지(阻止)시켰고 무처리구(無處理區)보다 더욱 효과적(效果的) 으로 Adenine 요구변이주(要求變異株)를 약(約) 6배정도(培程度) 더 분리(分離)케 하였다. E. coli에 대(對)하여도 위의 최적조건(最適條件)으로 조사(照射)하는데 Streptomycin$(200\;{\mu}g/ml)$ 처리구(處理區)는 생육(生育)을 현저(顯著)히 조지(阻止)시켰고 무처리구(無處理區)보다 Adenine요구변이주(要求變異株)를 2배(倍)나 효과적(效果的)으로 분리(分離)할 수 있었다.

• 미생물학회지
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• 제24권2호
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• pp.91-97
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• 1986

Intra and interspecfic fusants were produced by the protoplast fusion of auxotrophic mutants from Trichoderma koningii ATCC 26113 and Trichoderma reesei QM 9414. It was found that 0.6M $MgSO_4\;and\;0.6M\;NH_4Cl$ was the best osmotic stabilizer for the preparation of protoplasts from the mycelium of T. koningii and T. reesei respectively. However, $MgSO_4$ was the most suitable one for the regeneration of the protoplasts from both species. The intraspecific protoplast fusion frequencies between the auxotrophic mutants from T. reesei were $1.8{\times}10^{-2}\;to\;5.1{\times}10^{-1}$. Interspecific protoplast fusion frequencies between the auxotrophic mutants from T. koningii and T. reesei were $3.6{\times}10^{-3}$\;to\;8.4{\times}10^{-2}. Interspecific complementing fusants, however, were not alwats produced. Fusants obtained from interspecific potoplast fusion were spontaneously segregated into various strains including parental types, non-parental auxotrophic hybrids, and prototrophic hybrids on complete plate. Interspecific hybrids revealed to have partially enhanced celluloytic activities.

• 한국균학회지
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• 제25권1호통권80호
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• pp.26-29
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• 1997

여름느타리버섯의 담자포자에 자외선을 조사하여 5'-fluoro-orotic acid(FOA) 선발배지에서 pyrimidine 영양요구성 균주를 직접 선발하였다. 24개의 5'-FOA 저항성 균주중 13개의 pyrimidine 영양요구성 균주를 선발할 수 있었으며, 이들의 mating type group과 성장속도를 측정하여 형질전환을 위한 최적의 DNA수용체를 선발할 수 있었다.

• 미생물학회지
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• 제22권2호
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• pp.103-110
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• 1984

The conditions for the protoplast fusion of auxotrophic mutants of Trichoderma koningii were determined. A preparation of commercial enzyme Driselase was used successfully to isolate protoplasts from the 18 hr old mycelium of T. koningii. The yields of protoplasts production were ranged from $0.3{\times}10^8$ to $2.5{\times}10^8$ protoplasts per mg of damp mycelium of various auxotrophic mutant strains. The regeneration frequencies from $9.3{\times}10^{-3}\;to\;2.0{\times}10^{-1}$ were obtained when the protoplasts from auxotrophic mutants were plated on the malt extract medium containing 0.6M $MgSO_4$, and 2% agar, and the optimal concentration of PEG for protoplst fusion was 30%. Exposure of protoplasts to PEG for 10 min was found to be sufficient to induce high frequency heterokaryon formation. Optimal pH of fusion mixture was determined as 5.5, and 1 mM of calcium chloride in fusion mixture was found to be sufficient to enhance protoplast fusion frequency. Under optimal condition, the fusion frequency of the cross between protoplasts from various auxotrophic mutants were $1.6{\times}10^{-2}\;and\;4.1{\times}10^{-2}$.

• 미생물학회지
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• 제20권1호
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• pp.1-10
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• 1982

From Aspergillus nidulans, six suppressor mutants, MSI-MS6, were isolated, and their characteristics were analysed. These were the suppressor mutants for acriflavin resistant and nicotinamide auxotrophic mutant phenotypes. MS1, MS2, MS5 and MS6, were linked to the chromosome IV, I, II respectively, and MS2 was linked to one of the rest chromosomes, MS3 and MS6 mutants had both suppressors for acriflavin resistant marker and for nicotinamide auxotrophic marker. In order to know the stability and efficiency of the suppressors, their reversion frequencies, that is, frequencies of losing the suppressibility, were analysed. Only MS3 and MS5 were reversed with high frequency. The four mutants didn't lose their suppressibilities, and this meant that the suppressors of these four were very stable and highly effcient. The suppressor specificities of these mutants were tested for other mutant's phenotype marker. One of the six suppressors, MS1, had the suppressor specificity for acriflavin resistant marker of 163 strain.

• 한국균학회지
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• 제21권3호
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• pp.195-199
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• 1993

Fusarium oxysporum을 재료로 하여 polygalacturonase 활성능이 좋은 우량 균주를 개발하기 위하여 돌연변이주와 원형질체융합주를 선별하였다. MNNG를 처리하여 6개의 영양요구성 돌연변이주를 유도하였다. 세포벽 분해효소로는 driselase를 이용하고 삼투안정제로 0.6M KCI을 처리하여 돌연변이체로부터 원형질체를 얻었다. polyethyleneglycol 8,000과 $CaCl_2$를 이용하여 영양요구성 돌연변이체의 원형질체 사이에 융합을 시도하여 1개의 융합주를 얻었으며 융합률은 $5.0{\times}10^{-3}$이었다. polygalacturonase 활성도를 수정된 DNS법을 이용하여 측정하였다. 결과적으로, 선별된 융합주와 돌연변이주의 polygalacturonase 활성은 모균주와 비교하여 약 1.5배에서 3.5배가량 증가하였으며 그 중에서도 mutant Fx-2가 가장 높은 활성도를 나타냈다. 그러므로 이 방법은 우량균주개발을 위한 유용한 방법으로 판단되었다.

• 한국균학회지
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• 제14권2호
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• pp.185-188
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• 1986

노랑 느타리버섯의 유전 연구 및 다른 느타리종과의 융합을 위해서 노랑 느타리의 담자포자에 자외선을 조사하여 영양요구성 균주를 선발하였다. 노랑 느타리 포자에 UV를 조사한 결과 UV조사시간이 $40{\sim}70$초이고 생존율 $0.4{\sim}1.4%$ 일때 많은 mutants를 얻을 수 있었고, 선발된 14 auxotrophic mutants의 유전 marker는 amino acid-requiring strain 3균주, nucleic acid base-requiring strain 1 균주, vitamin-requiring strain 3 균주, amino acid/nucleic acid base-requiring strain 2 균주, amino acid/vitamin-requiring strain 5 균주로 되었다.

• 한국균학회지
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• 제13권2호
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• pp.75-78
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• 1985

진균류의 균주개발과 유전연구에 필수적인 영양요구성 균주 선발을 위하여 느타리비섯과 사철느타리버섯의 월형질체에 자외선을 조사하여 얻은 결과를 요약하면 다음과 같다. 1. 원형 질체에 자외선을 조사하여 그 생존을이$0.83{\sim}15%$일 때 가장 많은 영양 요구성 균주를 선발하였다. 2. 느타리에서 Arg, Gly Ser 그리고 Ala Orn Tyr 3균주와 사철느타리버섯에서 Ribo-1, Ribo-2, Phen, 그리고, Ade Hyp 4군주를 선발하였으며 이들 중에서 Phen 균주는 back mutation 현상이 나타났다.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1994년도 한국생물과학협회 학술발표대회
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• pp.251.2-251
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• 1994

No Abstract, See Full Text

• 미생물학회지
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• 제29권2호
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• pp.123-129
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• 1991

In order to develop the protoplast fusion method of the strains of Fusarium, the interspecific protoplast fusion was attempted between Fusarium poae and F. sporotrichioides. Various auxotrophic mutants were isolated by the treatment of N-Methyl-N'-Nitro-N-Nitrosoguanidine. The optimal conditions for the formation and regeneration of protoplasts were examined and the characteristics of a fusant were studied. As a results, protoplasts were readily obtained from 18 hours cultured mycelia by the treatment of driselase for 3 hours and 0.6 M KCl as a best osmotic stabilizer at pH 6.0 for the formation of protoplast. Sucrose was the most suitable for the regeneration. Polyetylene glycol (M.W. 8,000) in $CaCl_{2}$-glycine solution was used to induce the protoplast fusion. The interspecific fusion frequency between protoplasts among the auxotrophic mutants of the two strains ranged from $2.7*10^{-2}$ to $5.7*10^{-3}$ . DNA content and cellulase activity were rather increased in the interspecific fusant. The lag phase of growth curve was slightly elongated in the fusant.