中文摘要：本发明涉及一种用于转基因小麦检测的标准质粒分子及其构建方法，其特征是利用重叠PCR和分子克隆的方法构建了含有小麦内参基因WaXY?D1、转基因小麦常见转基因元件Ubiquitin、NPTII、Bar、T?NOS及品系B73?6?1特异性序列的标准质粒分子，并通过验证实验证明此标准分子可作为阳性标准物质用于转基因小麦的PCR检测，很好的解决了转基因小麦检测过程中阳性标准物质缺乏的问题。
外文摘要：NOVELTY - A standard plasmid molecule and its construction method, are new. The standard plasmid molecule comprises wheat internal reference gene WaXY-D1 fragment, transgenic element ubiquitin, transgenic element neomycin phosphotransferase (NPTII), transgenic element Bar gene, transgenic element nopaline synthase terminator (T-NOS) fragment, wheat transgenic line B73-6-1 specific sequence, in series.
USE - The standard plasmid molecule is used as positive reference substance for detecting transgenic wheat (all claimed) by PCR.
TF   TECHNOLOGY FOCUS - BIOTECHNOLOGY - A standard plasmid molecule and its construction method, are new. The standard plasmid molecule comprises wheat internal reference gene WaXY-D1 fragment, transgenic element ubiquitin, transgenic element neomycin phosphotransferase (NPTII), transgenic element Bar gene, transgenic element nopaline synthase terminator (T-NOS) fragment, wheat transgenic line B73-6-1 specific sequence, in series, where Pst I and HindIII cleavage sites are found in B73-6-1 specific sequence of wheat transgenic line, and the transgenic line B73-6-1 specific sequence in series comprises 3713 base pair sequence (SEQ ID No: 1) fully defined in the specification. Preferred Biomolecule: The wheat internal reference gene WaXY-D1 is wheat starch synthase gene, where the wheat starch synthase gene is highly conserved in wheat genome and has characteristics of interspecificity, intra-nonspecific and single copy number. The wheat transgenic element ubiquitin is maize ubiquitin gene promoter or transgenic monocotyledons exogenous gene promoter. The transgenic element NPTII is neomycin-3'-phosphotransferase gene for antibiotic screening in transgenic plants. The transgenic element Bar is glyphosate acetyltransferase gene for enhancing resistance of transgenic plants to herbicide glufosinate. The wheat transgenic element T-NOS is nopaline synthase terminator as terminator for exogenous gene of transgenic plants. The wheat strain B73-6-1 specific sequence is wheat transgenic line B73-6-1 exogenous DNA sequence inserted into vector.
主权项：一种用于转基因小麦检测的标准质粒分子及其构建方法，其特征在于小麦内参基因WaXY?D1片段、转基因元件Ubiquitin、NPTII、Bar、T?NOS片段、小麦转基因品系B73?6?1特异性序列依次串联，在小麦转基因品系B73?6?1特异性序列后还有Pst?I和HindIII酶切位点，该串联序列的序列如SEQ?ID?No：1所示。
申请号：CN201610396777.3
专利号：CN106244674-A
公开/公告号：CN106244674-A
申请/专利权人：谱尼测试集团深圳有限公司
发明/设计人：宋薇;张斌;崔山;郭文宗
分类号：C12Q1/68;C12N15/11;C12N15/10
主分类号：C12Q1/68
法律状态：公开专利
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