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    • 30. 发明申请
    • SCRAMBLE OF HETEROZYGOUS DIPLOID YEAST
    • 杂合双杂体酵母的混合物
    • WO2018009863A1
    • 2018-01-11
    • PCT/US2017/041194
    • 2017-07-07
    • NEW YORK UNIVERSITYTIANJIN UNIVERSITY
    • SHEN, MichaelBOEKE, Jef, D.WU, YiYUAN, Yingjin
    • C12N1/36C12N15/10C12N15/52C12N15/64C12N15/66C12N15/81
    • Provided are compositions and methods for making a diploid yeast having heterozygosity for at least one chromosome. The method includes mating haploid yeast having at least a first modified chromosome comprising a synthetic chromosome suitable for recombination-site-mediated evolution (as a SCRaMbLE-ready modification). The SCRaMbLE-ready modification includes introduced site-specific recombinase recognition sites that can be recognized by a recombinase. Yeast that have at least one SCRaMbLE-ready modification of a chromosome are mated with a haploid yeast devoid of the SCRaMbLE- ready modification to obtain diploid SCRaMbLE-ready yeast. Subsequent to mating the haploid SCRaMbLE-ready yeast to yeast devoid of the SCRaMbLE-ready modification the method includes using the recombinase to recombine the first modified chromosome to obtain heterozygous diploid yeast comprising at least one recombined (SCRaMbLEd) chromosome and a homologous non-SCRaMbLEd chromosome. The method further includes identifying heterozygous diploid yeast that comprise the at least one SCRaMbLEd chromosome that have a changed phenotype that is different from the phenotype of the diploid SCRaMbLE-ready yeast. Yeast made by the methods, compositions of matter made by the yeast, spores made by the yeast, and kits containing the yeast and/or their spores are included.
    • 提供了用于制造具有至少一个染色体的杂合性的二倍体酵母的组合物和方法。 该方法包括交配具有至少第一修饰染色体的单倍体酵母,所述第一修饰染色体包含适合于重组位点介导的进化的合成染色体(作为SCRaMbLE就绪修饰)。 SCRaMbLE就绪修饰包括引入的位点特异性重组酶识别位点,可以被重组酶识别。 对至少有一个SCRaMbLE就绪修饰染色体的酵母与没有SCRaMbLE-ready修饰的单倍体酵母交配以获得二倍体SCRaMbLE准备好的酵母。 在将单倍体SCRaMbLE就绪酵母与不含SCRaMbLE就绪修饰的酵母交配之后,所述方法包括使用重组酶将第一修饰染色体重组以获得杂合二倍体酵母,所述杂合二倍体酵母包含至少一个重组(SCRaMbLEd)染色体和同源非SCRaMbLEd 染色体。 该方法进一步包括鉴定杂合二倍体酵母,该杂合二倍体酵母包含至少一个SCRaMbLEd染色体,其具有改变的表型,该表型与二倍体SCRaMbLE就绪型酵母的表型不同。 包括由该方法制造的酵母,由酵母制成的物质组成,由酵母制成的孢子和含有酵母和/或其孢子的试剂盒。