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Method for instantly expressing by converting exogenous gene into lotus

A technology of exogenous gene and instantaneous expression, applied in the field of instantaneous expression, can solve the problems of immature genetic transformation system, difficult regeneration of explants, slow progress in lotus transgenic research, etc., and achieve easy implementation, simple operation method and short time Effect

Inactive Publication Date: 2016-12-14
SHANGHAI CHENSHAN BOTANICAL GARDEN
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main reason is that the genetic transformation and explant regeneration of lotus has always been difficult. So far, there have been few successful reports, and the genetic transformation system is still very immature, resulting in very slow progress in the transgenic research of lotus.

Method used

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  • Method for instantly expressing by converting exogenous gene into lotus
  • Method for instantly expressing by converting exogenous gene into lotus
  • Method for instantly expressing by converting exogenous gene into lotus

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Experimental program
Comparison scheme
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Embodiment

[0041] A method for transiently expressing exogenous genes into lotus flowers, comprising the steps of:

[0042] 1) Gather flower buds

[0043] Select unexpanded ‘M283’ lotus flower (large, single-petal red flower) buds (see figure 1 ), 'L097' lotus (large and medium-sized, white flowers with double petals) buds or 'American yellow lotus', the length of the buds is > 5cm, and the length of the flower stalk is 10-20cm. Put the flower stalk in clean water and cultivate it indoors , the temperature is 25~30℃;

[0044] 2) Prepare positive Agrobacterium

[0045] Transform the pCAMBIA 1301 plasmid carrying the GUS gene into the plant Agrobacterium GV 3101 and LBA 4404 respectively, and use the petals of the GV 3101 Agrobacterium without the GUS gene as a control, pick a single clone, and obtain positive Agrobacterium by PCR electrophoresis. ;

[0046] 3) Preparation of bacterial liquid

[0047] Pick the monoclonal positive Agrobacterium, transfer it to YEB liquid medium, shake ...

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Abstract

The invention relates to a method for instantly expressing by converting exogenous gene into lotus. The method mainly comprises the steps of collecting flower buds; preparing positive agrobacterium; preparing bacterium solution; converting the exogenous gene and culturing; detecting the instant expression; converting the expression plasmid containing the exogenous gene into the tissues of the lotus petals by adopting the agrobacterium according an injection penetrating method; culturing, thereby instantly expressing the exogenous gene in the petals. According to the invention, the exogenous gene is converted into the lotus petals for instantly expressing, so that the petal tissues are not damaged; the agrobacterium is easy to diffuse; after the exogenous gene is converted, a test result can be observed after 40-48 hours; the operation method is simple and convenient; the converting rate is high; the required time is short; the exogenous gene can be quickly detected; the verification for the functional gene of the lotus can be effectively performed.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a method for transforming exogenous genes into lotus flowers for instant expression. Background technique [0002] The lotus is a perennial aquatic herb. It is the national flower of India and a world-famous religious sacred flower. It is widely distributed in my country and has a long history of cultivation. It is one of the top ten traditional Chinese flowers and has extremely high cultural connotation and ornamental value. [0003] Lotus not only has profound cultural heritage, but also has extremely high economic value. Lotus root is one of the important vegetables in our country. Lotus seeds and lotus leaves have health effects such as regulating blood lipids. As a well-known ornamental plant, lotus can be used as potted or cut flowers. It can not only beautify the environment, but also purify water. Therefore, lotus is a very important flower that integrates ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
Inventor 田代科张大生陈青刘青青刘凤栾赵喜双吴璇陈蒙娇吴少华
Owner SHANGHAI CHENSHAN BOTANICAL GARDEN
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