Beta-Glucosidase 1 (bgl1) Gene Analysis in Mutant and Wild-type of Penicillium sp. ID10-T065

Vilya Syafriana, S. Nuswantara, W. Mangunwardoyo, P. Lisdiyanti
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Abstract

In the previous study, Penicillium sp. ID10-T065 has been mutated using ultraviolet (UV), ethyl methyl sulfonate (EMS) and the combination of UV-EMS to increase Beta-glucosidase (bgl) activity. There were three mutants selected, UV13 (UV mutant), EM31 (EMS mutant), and UM23 (UV-EMS mutant). This study examined the mutations in the bgl gene encoding (bgl1) as well as sequence differences between mutants and wild-type of Penicillium sp. ID10-T065. The gene analysis was performed by PCR amplification and sequencing of the bgl1 gene. The results of bgl1 gene sequences (600 bp) from mutants were aligned with the wild-type, it was discovered that there were base alterations from position 2025 to 2050. Mutant UV13 showed the highest base alterations (7 bases) which occurred at position 2027 (T→C); 2036 (T→G); 2040 (T→G); 2047 (G→C); and 2048-2050 (TTG→GGA). Mutant EM31 showed alterations in five bases at positions 2034 (G→A), 2036 (T→G), 2037 (G→C), 2044 (G→C), and 2047 (G→T). Mutant UM23 showed two base alterations at position 2025 (T→A) and 2037 (G→C). UV irradiation and EMS mutation resulted in transition and transversion DNA, whereas the combination of UV-EMS mutation resulted in transversion mutations. Base alterations in UV13 and EM31 mutants, causing missense and silent mutation, while in UM23 mutant only silent mutations occur. The bgl1 gene analysis showed that mutation using UV light was more effective than using EMS or a combination of UV-EMS in Penicillium sp. ID10-T065.
青霉菌ID10-T065突变型与野生型β -葡萄糖苷酶1基因分析
在先前的研究中,利用紫外线(UV)、甲基磺酸乙酯(EMS)和紫外-EMS联合诱变青霉sp. ID10-T065,以提高β -葡萄糖苷酶(bgl)的活性。选择了三个突变体,分别是UV13 (UV突变体)、EM31 (EMS突变体)和UM23 (UV-EMS突变体)。本研究检测了青霉菌ID10-T065的bgl基因编码(bgl1)突变以及突变体与野生型之间的序列差异。对bgl1基因进行PCR扩增和测序分析。突变体bgl1基因序列(600 bp)与野生型一致,发现2025 ~ 2050位点存在碱基变化。突变体UV13的碱基变化最多(7个碱基),发生在2027位点(T→C);2036 (T→G);2040 (T→G);2047 (G→C);2048 ~ 2050年(TTG→GGA)。突变体EM31在2034 (G→A)、2036 (T→G)、2037 (G→C)、2044 (G→C)和2047 (G→T)位点上有5个碱基的改变。突变体UM23在2025位点(T→A)和2037位点(G→C)有两个碱基改变。紫外辐照和EMS突变导致DNA的过渡和翻转,而紫外-EMS联合突变导致DNA的翻转突变。在UV13和EM31突变体中发生碱基改变,导致误义和沉默突变,而在UM23突变体中只发生沉默突变。对ID10-T065青霉bgl1基因的分析表明,紫外诱变比紫外诱变或紫外-EMS联合诱变更有效。
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