Positive plasmid for identification of transgenic rice transformants and its construction method and application

A technology of transgenic rice and positive plasmids, which is applied in chemical instruments and methods, biochemical equipment and methods, and introduction of foreign genetic material using vectors, can solve the problems of narrow coverage, poor applicability and specificity, etc. The effect of facilitating popularization and application, reducing labor cost and economic cost

Active Publication Date: 2020-12-22
RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI
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  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

These public multi-target positive plasmid molecules can only cover three transgenic lines, and have defects such as narrow coverage, poor applicability and specificity, and need to be improved

Method used

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  • Positive plasmid for identification of transgenic rice transformants and its construction method and application
  • Positive plasmid for identification of transgenic rice transformants and its construction method and application
  • Positive plasmid for identification of transgenic rice transformants and its construction method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Construction, extraction, purification and sequencing verification of positive plasmid molecules

[0044] 1. Determine the detection target and sequence of each rice transformant

[0045] 11 rice transformants were obtained by searching the "GM ApprovalDatabase" database of the International Agricultural Biotechnology Application Service Organization (ISAAA) website, related patents, literature, and isolation, identification, and sequencing to obtain the exogenous expression cassette inserted by the exogenous gene in the rice transformation. and the nucleotide sequence information of 2 standard genes in rice:

[0046] 11 rice transformants: TT51-1 / Huahui 1 / Shanyou 63, T1C-19, T2A-1, Kelangdao 1, Kefeng 2, Kefeng 6, Kefeng 8, G6H1, M12 , PA110-15, 114-7-2;

[0047] 2 rice internal standard genes: SPS (2 fragments) and PLD.

[0048] The detection primer sequences and detection methods of each rice transformant were obtained through the transgenic detection me...

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Abstract

The invention discloses a positive plasmid for identifying a transgenic rice transformant as well as a construction method and an application thereof. The positive plasmid comprises a vector skeletonand a screening element, the screening element is composed of 11 rice transformant flanking sequences and two rice internal standard genes, and the two rice internal standard genes are respectively anSPS gene and a PLD gene. The positive plasmid constructed by the invention can be applied to screening and detection of transgenic rice; the method comprises the following steps: taking the positiveplasmid as a positive control plasmid or a positive standard substance for transgenic rice detection; and carrying out common PCR or real-time fluorescent PCR screening detection on transgenic components of a rice sample to-be-detected; the positive plasmid can accurately detect various transgenic rice, meets the requirements of safety supervision and detection work of the transgenic rice, avoidsthe problem that a plurality of positive controls need to be prepared for a plurality of detection targets in the existing detection and identification process, and has the characteristics of wide coverage area, easiness in obtaining a large amount of positive controls, simplicity in preparation and the like.

Description

technical field [0001] The present invention relates to a positive plasmid for identification of transgenic plant transformants, in particular to a positive plasmid for identification of transgenic rice transformants and a construction method thereof. The present invention further relates to the application of the positive plasmid in identifying 11 kinds of transgenic rice transformants, belonging to transgenic Field of identification of rice. Background technique [0002] Rice, as the staple food of more than half of the world's population, is one of the important food crops. As an emerging breeding method, transgenic technology has been widely used in crop quality improvement. So far, many studies on transgenic rice with insect resistance, disease resistance, and herbicide tolerance have been reported. In China, currently only the transgenic insect-resistant rice variety Huahui 1 and its hybrid Bt Shanyou 63 have obtained the production and application safety certificate...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12Q1/6895
CPCC07K14/415C12Q1/6895C12Q2600/158
Inventor 马卉张秀杰汪秀峰焦小雨许学李夏莹吴爽陈子言王颢潜梁晋刚
Owner RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI
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