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Question

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Two different phenotypic classes of recombinant strains can be generated Mut+ and MutS refers to the “Methanol utilization slow” phenotype caused by the loss of alcohol oxidase activity encoded by the AOX1 gene. A strain with a MutS phenotype has a mutant aox1 locus, but is wild type for AOX2. This results in a slow growth phenotype on methanol medium. Both X-33 and GS115 are Mut+, and KM71H is MutS. Transformation of X-33 or GS115 with plasmid DNA linearized in the 5’ AOX1 region will yield Mut+ transformants, while KM71H will yield only MutS transformants. Both Mut+ and MutS recombinants are useful to have one phenotype may favor better expression of your protein than the other. You should test between 6-10 recombinants per phenotype because the site of recombination may affect expression. There is no way to predict beforehand witch construct or isolate will better express your protein.

Answer 1

關於嗜甲醇酵母菌菌珠介紹
原文：
Two different phenotypic classes of recombinant strains can be generated Mut+ and MutS refers to the "Methanol utilization slow" phenotype caused by the loss of alcohol oxidase activity encoded by the AOX1 gene. A strain with a MutS phenotype has a mutant aox1 locus, but is wild type for AOX2. This results in a slow growth phenotype on methanol medium. Both X-33 and GS115 are Mut+, and KM71H is MutS. Transformation of X-33 or GS115 with plasmid DNA linearized in the 5' AOX1 region will yield Mut+ transformants, while KM71H will yield only MutS transformants. Both Mut+ and MutS recombinants are useful to have one phenotype may favor better expression of your protein than the other. You should test between 6-10 recombinants per phenotype because the site of recombination may affect expression. There is no way to predict beforehand witch construct or isolate will better express your protein.

譯文：
可產生有兩種顯性基因重組的種系，即為Mut+和MutS得稱為「乙醇使用效率緩慢」的基因重組的種系，該種系是缺乏可由AOXI 基因所引發的酒精氧化酵素活動所造成的。某種含有MutS顯性基因重組的種系，即含有突變的AOX1基因座，但係屬AOX2的突變型。本結果即會在乙醇的培養基中，緩慢成長形成顯性基因重組的種系。 X-33 和 GS115即為Mut+，而KM71H即為MutS。在5' AOX1的區域中，出現有線性化質體DNA的X-33 或GS115的轉化現象，即會產生Mut+的轉化體，而KM71H只會產生MutS的轉化體。Mut+和MutS的轉化體得用於讓顯性基因重組種系比起其他的種系而言，會在蛋白質上有更佳的表現。可在各顯性基因重組的種系中，測試6-10個轉化體，因為轉化體的區域可能會影響基因的表現。無法事先預測其可能出現的不良結構或加以隔離，而讓蛋白質有更佳的表現。

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