首页> 外国专利> AMYLOPECTIN POTATO STARCH WITH IMPROVED STABILITY AGAINST RETROGRADATION AND IMPROVED FREEZE AND THAW STABILITY

AMYLOPECTIN POTATO STARCH WITH IMPROVED STABILITY AGAINST RETROGRADATION AND IMPROVED FREEZE AND THAW STABILITY

机译:淀粉变性的淀粉淀粉,具有改进的抗逆稳定性和改善的冻融稳定性

摘要

Amylopectin potato starch with improved stability against retro-gradation and improved freeze and thaw stability, wherein it contains more than 99% amylopectin, preferably 100% amylopectin, is disclosed, as well as a method for the production of a potato (Solanum tuberosum) containing said amylopectin potato starch, wherein said method involves homology-directed mutagenesis using CRISPR/nuclease technology and comprises the following steps: a) provision of potato cells or potato tissue containing potato cells, b) introduction into the nuclei of said potato cells of one or more CRISPR/nuclease complexes each comprising a specific targeting ribonucleotide sequence which is fully or essentially homologous to a target nucleotide sequence located in a DNA sequence immediately upstream of a PAM (5′-NGG-3′protospacer adjacent motif) in a gene coding for a GBSS enzyme and optionally also in a gene coding for an SSII enzyme and/or in a gene coding for an SSIII enzyme, wherein said mutagenesis takes place in one or more alleles of the potato genome, wherein when said targeting ribonucleotide sequence identifies the complementary strand of the target nucleotide sequence, said one or more CRISPR/nuclease complexes cut(s) said DNA sequence, leading to a subsequent complete lack of the ability of the potato to produce a functional GBSSI enzyme, optionally also a functional SSII and/or SSIII enzyme, c) wherein step b) optionally is repeated until the potato lacks the ability to produce said functional GBSSI enzyme, optionally also a functional SSII and/or SSIII enzyme, in all of the alleles, preferably 3 times, a potato obtained by said method, a method for the production of said amylopection potato starch from said potato, and different uses of said amylopectin potato starch. a gene coding for an SSII enzyme and/or in a gene coding lor an SSIII enzyme, wherein said mutagenesis lakes place in one or more alleles of the potato genome, wherein when said targeting ribonucleotide sequence identifies the complementary strand of the target nucleotide sequence, said one or more CRISPR/nuclease complexes cut(s) said DNA sequence, leading to a subsequent complete lack of the ability of the potato to produce a functional GBSSI enzyme, optionally also a functional SSII and/or SSIII enzyme, c) wherein step b) optionally is repeated until the potato lacks the ability to produce said functional GBSSI enzyme, optionally also a functional SSII and/or SSIII enzyme, in all of the alleles, preferably 3 times, a potato obtained by said method, a method for the production of said amylopection potato starch from said potato, and different uses of said amylopectin potato starch.
机译:公开了一种支链淀粉马铃薯淀粉,其具有改善的逆凝稳定性和改善的冷冻和融化稳定性,其中包含超过99%的支链淀粉,优选100%的支链淀粉,以及一种生产含马铃薯的马铃薯(Solanum tuberosum)的方法。所述支链淀粉马铃薯淀粉,其中所述方法包括使用CRISPR /核酸酶技术的同源性定向诱变,并且包括以下步骤:a)提供马铃薯细胞或含有马铃薯细胞的马铃薯组织,b)将一种或多种马铃薯细胞核引入所述马铃薯细胞核中。更多CRISPR /核酸酶复合物,每个复合物均包含特定的靶向核糖核苷酸序列,该序列与编码PAM的PAM(5'-NGG-3'protospacer相邻基序)上游的DNA序列中的目标核苷酸序列完全或基本同源。 GBSS酶,也可以选择在编码SSII酶的基因中和/或在编码SSIII酶的基因中,其中所述诱变在马铃薯基因组的一个或多个等位基因中发生,其中当所述靶向核糖核苷酸序列识别靶核苷酸序列的互补链时,所述一个或多个CRISPR /核酸酶复合物切割所述DNA序列,从而导致随后的完全缺失产生功能性GBSSI酶,任选地还有功能性SSII和/或SSIII酶的能力的测定,c),其中步骤b)任选地重复,直到马铃薯缺乏产生所述功能性GBSSI酶,任选地还有功能性的能力在所有等位基因中,SSII和/或SSIII酶(优选3倍)是通过所述方法获得的马铃薯,从所述马铃薯生产所述支链淀粉马铃薯淀粉的方法以及所述支链淀粉马铃薯淀粉的不同用途。编码SSII酶的基因和/或编码SSIII酶的基因,其中所述诱变湖位于马铃薯​​基因组的一个或多个等位基因中,其中当所述靶向核糖核苷酸序列确定靶核苷酸序列的互补链时,所述一种或多种CRISPR /核酸酶复合物切割所述DNA序列,导致马铃薯随后完全缺乏产生功能性GBSSI酶,任选地还有功能性SSII和/或SSIII酶的能力,c),其中步骤b)任选地重复进行,直到马铃薯在所有等位基因中缺乏产生所述功能性GBSSI酶,任选地还缺乏功能性SSII和/或SSIII酶的能力,优选为通过所述方法获得的马铃薯的3倍。由所述马铃薯生产所述支链淀粉马铃薯淀粉,以及所述支链淀粉马铃薯淀粉的不同用途。

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