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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 biogenetic engineering

Inactive Publication Date: 2014-08-27
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, salt stress can also induce some plants to produce amino acid transporters. For example, salt stress induced Arabidopsis to produce two proline transporters (Rentsch, 1996), but Igarashi et al. (2000) reported that proline in rice plants The transporter (OsProT) is not induced by salt stress

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
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Effect test

Embodiment 1

[0023] Implementation Example 1 The acquisition of PgLHT gene

[0024] 1. Ginseng suspension cell culture

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

[0026] (2) Under sterile conditions, the sterilized explants were inoculated 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. Subculture to supplement the nutrients and other requirements of the culture. The first subculture was carried out 30 days after inoculation, and then every 3-4 w...

Embodiment 2

[0086] Implementation example 2 construction and transformation of expression vector

[0087] 1. Construction of plant overexpression vectors and screening of transgenic plants

[0088] (1) Digest pBI121 and PgLHT with BamH I and Sac I respectively, and recover pBI121 vector and PgLHT gene fragment. The two fragments were connected and transformed into DH5α, and the recombinant was screened out, and the recombinant plasmid was named

[0089] pBI121-PgLHT. The upstream and downstream primers for amplifying the PgLHT gene are respectively:

[0090] PgLHT: F1:5′-CG GGATCC CGCACCATGGTTTCCCAAAGCTCAGAC-3'; (SEQ ID NO. 11)

[0091] R1:5′-CGC GAGCTC GGCTCCTTTGCTGCTTATGAGAA-3'. (SEQ ID NO.12)

[0092] The underlined part is the restriction site of BamH I and Sac I.

[0093] (2) Preparation of Agrobacterium tumefaciens Competent Cells

[0094] ① Streak culture of Agrobacterium LBA4404 on YEB plate containing 100 μg / ml rifamycin, culture in dark at 28°C for 2 days.

[0095] ②...

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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 biogenetic engineering, and relates to improving plant traits, in particular to related genes, proteins and applications thereof that promote amino acid translocation and participate in plant stress responses such as ginseng. The invention has important application value in regulating plant organic nutrition metabolism, defense response and improving cultivation quality of ginseng and other plants. Background technique [0002] Ginseng (P.ginseng C.A.Meyer) is a perennial herb of the Araliaceae genus Panax, and is a precious Chinese medicinal material. However, ginseng is a plant that avoids continuous cropping, and 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 cultivation and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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