中文摘要：本发明公开了一种大豆转化事件ZUTS?33，所述转化事件ZUTS?33中含有一个外源插入序列，所述外源插入序列的5'端与内源基因组DNA构成的连接序列如SEQ?ID?NO:1所示，所述外源插入序列的3'端与内源基因组DNA构成的连接序列如SEQ?ID?NO:2所示。本发明还同时公开了相应的用于鉴定转基因大豆ZUTS?33的检测方法、检测生物样品中事件ZUTS?33或其子代的方法、检测事件ZUTS?33中外源插入DNA分子的方法、检测来源于ZUTS?33事件的植物材料的方法等内容。
外文摘要：NOVELTY - A transgenic soybean ZUTS-33 comprising externally inserted sequence, where the externally inserted sequence at 5' end of endogenous genomic DNA comprises a 359 base pair sequence (SEQ ID NO: 1) fully defined in the specification, and the externally inserted sequence at 3' end of endogenous genomic DNA comprises a 460 base pair sequence (SEQ ID NO: 2) fully defined in the specification, is new.
USE - As transgenic soybean ZUTS-33.
DETAILED DESCRIPTION - INDEPENDENT CLAIMS are included for the following:
(1) method (M1) for identifying transgenic soybean ZUTS-33, involves (a) designing specific sequence for specifically identifying 5' end of T-DNA insertion or first primer comprising base pair sequence of SEQ ID NO: 4 (identifying the primers on 5' end of genome), and base pair sequence of SEQ ID NO: 5 (identifying primers for T-DNA insertion sequence), or its complementary sequence, and (b) designing specific sequence for specifically identifying 3' end of T-DNA insertion or first primer comprising base pair sequence of SEQ ID NO: 6 (identifying the primers on 3' end of genome), and base pair sequence of SEQ ID NO: 7 (identifying primers for T-DNA insertion sequence), or its complementary sequence;
(2) method (M2) for detecting transgenic soybean ZUTS-33 or its progeny in a biological sample, involves (a) extracting a DNA sample from the biological sample, (b) providing DNA primer pairs, (c) providing DNA amplification reaction conditions, (d) performing a DNA amplification reaction to obtain a DNA amplicon, (e) detecting DNA amplification product, where the DNA amplicon comprising 469 a 469 and 446 base pair sequence (SEQ ID NO: 8 and 9) fully defined in the specification;
(3) method (M3) for detecting externally inserted DNA molecules in transgenic soybean ZUTS-33;
(4) method (M4) for detecting a plant material derived transgenic soybean ZUTS-33;
(5) recombinant plasmid vector for soybean transformation compri! sing nuc leotide sequence functionally linked to at least one controlled expression polynucleotide, where the expression vector is pSOY19;
(6) method (M5) for constructing glyphosate-resistant transgenic soybean ZUTS-33;
(7) primer for detecting transgenic soybean ZUTS-33 comprising first pair of primer comprising base pair sequence of SEQ ID NO: 4 and 5, and the second pair of primers comprising base pair sequence of SEQ ID NO: 6 and 7; and
(8) nucleotide probe and an antibody for detecting transgenic soybean, where the nucleotide probe comprises a 1138 base pair or sequence (SEQ ID NO: 10) fully defined in the specification, the antibody specifically recognize gene expression of exogenous gene (g10-epsps) in transgenic soybean ZUTS-33, and the g10-epsps comprises a 1332 base pair sequence (SEQ ID NO: 11) fully defined in the specification. caaatgtaagtgggttgacagca (SEQ ID NO: 4), ccagtactaaaatccagatcccc (SEQ ID NO: 5), ggatttgaaattccgcctgac (SEQ ID NO: 6), and ctggcgtaatagcgaagagg (SEQ ID NO: 7).
主权项：大豆转化事件ZUTS?33，其特征是：所述转化事件ZUTS?33中含有一个外源插入序列，所述外源插入序列的5'端与内源基因组DNA构成的连接序列如SEQ?ID?NO:1所示，所述外源插入序列的3'端与内源基因组DNA构成的连接序列如SEQ?ID?NO:2所示。
申请号：CN201610825077.1
专利号：CN106498030-A
公开/公告号：CN106498030A
申请日：2016-09-18
公开/公告日：2017-03-15
申请/专利权人：浙江大学
发明/设计人：寿惠霞 ; 杜娟 ; 陆玲鸿 ; 李林 ; 宋张悦
分类号：C12Q1/68 ; C12N15/82 ; C12N15/11 ; C07K16/16 ; G01N33/68 ; G01N33/558 ; A01H5/00
主分类号：C12Q1/68
法律状态：实质审查
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