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Ginseng amino acid transport protein gene PgLHT and encoding protein and application thereof

A technology for transporting proteins and amino acids, applied in the field of biological genetic engineering, can solve the problems of nitrogen fertilizer ginseng quality and yield, environmental degradation, impact and other problems

Inactive Publication Date: 2013-03-06
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of ginseng cultivation and tissue and cell culture, inorganic nitrogen is usually the limiting factor for its growth. The unreasonable application of nitrogen fertilizer not only has a bad impact on the quality and yield of ginseng, but also causes environmental degradation.

Method used

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  • Ginseng amino acid transport protein gene PgLHT and encoding protein and application thereof
  • Ginseng amino acid transport protein gene PgLHT and encoding protein and application thereof
  • Ginseng amino acid transport protein gene PgLHT and encoding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Implementation Example 1 Acquisition of PgLHT Gene

[0024] 1. Ginseng suspension cell culture

[0025] (1) Take the four-year-old ginseng root from the ginseng production area of ​​Changbai Mountain in Jilin, soak it in tap water for 45 minutes, then rinse it twice, first soak it in 75% ethanol for 30 seconds (sec), rinse it twice with sterile water, and use it on the ultra-clean workbench in the sterile room. 0.1%HgCl 2 Sterilize, agitate constantly, rinse 4-5 times with sterile distilled water.

[0026] (2) Under sterile conditions, inoculate the sterilized explants into MS medium containing 2,4-D (2.0 mg / L) and KT (0.5 mg / L). After 3 subcultures, the explants were completely dedifferentiated. After the callus was completely formed, 6 explants were inoculated in each 100ml Erlenmeyer flask. The culture conditions were as described above. Subculture to supplement the nutrients and other requirements required by the culture. The first subculture was carried out 30 ...

Embodiment 2

[0081] Construction and transformation of embodiment 2 expression vector

[0082] 1. Construction of plant overexpression vector and screening of transgenic plants

[0083](1) pBI121 and PgLHT were digested with BamH I and Sac I, respectively, and the pBI121 vector and PgLHT gene fragments were recovered. The two fragments were ligated and transformed into DH5α, and the recombinant was screened out. The recombinant plasmid was named pBI121-PgLHT. The upstream and downstream primers for amplifying the PgLHT gene are:

[0084] PgLHT:F 1:5′-CG GGATCC CGCACCATGGTTTCCCAAGCTCAGAC-3′; (SEQ ID N0.11)

[0085] R1: 5′-CGC GAGCTC GGCTCCTTTGCTGCTTATGAGAA-3'. (SEQ ID NO.12)

[0086] The underlined parts are BamH I and Sac I restriction sites.

[0087] (2) Preparation of Agrobacterium tumefaciens competent cells

[0088] ①Agrobacterium LBA4404 was streaked on a YEB plate containing 100μg / ml rifamycin, and cultured in the dark at 28°C for 2 days.

[0089] ②Pick a single colony and...

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Abstract

The invention discloses a ginseng amino acid transport protein gene PgLHT and an encoding protein and an application thereof. The amino acid transport related gene PgLHT has a sequence as shown in SEQ ID NO.1, and the encoding protein has the sequence as shown in SEQ ID NO.2. Researches find that PgLHT genetic expression has the tissue specificity and is closely related to ginseng stress response. The PgLHT gene has obvious effect in enhancing the ginseng plant resistance and the like. Ginseng and other plants can be screened or improved through the gene or derivatives thereof, so that excellent plant varieties with strong environment adaptivity can be obtained. The organic nutrition metabolism of plants of ginseng and the like can also be promoted through adopting a tissue culture or transgene mode.

Description

technical field [0001] The invention belongs to the technical field of biological genetic engineering, and relates to improving plant traits, in particular to related genes, proteins and applications that promote amino acid transport and participate in plant stress responses such as ginseng. The invention has important application value in regulating plant organic nutrient metabolism, defense response and improving the cultivation quality of ginseng and other plants. Background technique [0002] Ginseng (P.ginseng C.A.Meyer) is a perennial herb of the Araliaceae ginseng genus and is a valuable Chinese medicinal material. However, ginseng is a plant that is not suitable for continuous cropping, and it has very strict requirements on soil nutrition, among which nitrogen is very important for the growth of ginseng. Many important compounds in ginseng contain nitrogen, such as proteins, nucleic acids, amino acids, pigments, and some growth hormones. In the process of ginseng ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/11C12N15/63C07K14/415A01H5/00
Inventor 罗志勇张儒祝捷谢小雷黄景嘉陈湘晖
Owner CENT SOUTH UNIV
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