A method for interfering with plant endogenous cle family polypeptide hormones and antagonistic polypeptides of small molecule polypeptide hormones

A technology for small molecule polypeptides and polypeptide hormones, which is applied in the field of genetic engineering and can solve problems such as functional research, functional redundancy, and difficulty in separating and detecting polypeptide hormones.

Active Publication Date: 2016-06-08
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many problems in the functional research of plant polypeptide hormones using the existing methods.
For example, 1: Since the genes encoding polypeptide hormones are usually small, current gene mutation methods such as EMS mutagenesis and T-DNA insertion methods are difficult to mutate them; 2: Since polypeptide hormones function as signal molecules in plants , the content is usually very low, and it is difficult to isolate and detect these polypeptide hormones from plants with current technical means; 3: Since the genes encoding plant polypeptide hormones usually belong to a certain gene family, the encoded precursor proteins contain conserved polypeptides The coding region has redundant functions. Even if one of the members is mutated, it is difficult to see that the plant is abnormal, so it is impossible to study its function

Method used

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  • A method for interfering with plant endogenous cle family polypeptide hormones and antagonistic polypeptides of small molecule polypeptide hormones
  • A method for interfering with plant endogenous cle family polypeptide hormones and antagonistic polypeptides of small molecule polypeptide hormones
  • A method for interfering with plant endogenous cle family polypeptide hormones and antagonistic polypeptides of small molecule polypeptide hormones

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Experimental program
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Embodiment 1

[0031] 1. One-dimensional alanine scanning of 12 amino acids of CLV3 polypeptide hormone

[0032] By designing primers and PCR, a point mutation is introduced into the CLE domain of the polypeptide hormone coding region of the CLV3 gene (sequence: N-RTVPSGPDPLHH-C), and the 12 amino acids in the polypeptide region are alanine (Alanine, A). Replacement, the PCR product (including 1857 bp before the start codon ATG at the 5' end and 1518 bp regulatory sequence and 559 bp of the genome sequence after the stop codon TGA at the 3' end, the nucleotide sequence of which is shown in SEQIDNo.1:) was cloned into pDONR221 Vector, and then subcloned into the pBGWFS7 destination vector, and the construction was named CLV3 in turn R1A ,CLV3 T2A ,CLV3 V3A ,CLV3 P4A ,CLV3 S5A ,CLV3 G6A ,CLV3 P7A ,CLV3 D8A ,CLV3 P9A ,CLV3 L10A ,CLV3 H11A and CLV3 H12A ( figure 1 ). These constructs were transformed into wild-type Arabidopsis thaliana by the method of Agrobacterium transformation c...

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Abstract

The invention relates to the field of genetic engineering and in particular relates to a method for interfering plant endogenous CLE family polypeptide hormone and antagonist polypeptide for small molecule polypeptide hormone. According to the method, effective antagonist polypeptides for small molecule polypeptide hormones of CLV3, CLE1, CLE8 and CLE27 are obtained through amino acid replacement and are expressed in plants through a transgenosis method, the specific polypeptides are subjected to functional interference, the downstream signal transduction is blocked, the polypeptides are further subjected to functional research, and the potential application values on model plant Arabidopsis and crops are mined.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for interfering with plant endogenous CLE family polypeptide hormones and an antagonistic polypeptide of small molecule polypeptide hormones. Background technique [0002] Since the discovery of systemin, the first polypeptide hormone in higher plants in the early 1990s, more than 20 small-molecule polypeptide hormones from different plants including Arabidopsis, rice, and soybean have been reported, involved in immune defense reactions , root nodule formation, cell division, stem cell maintenance and differentiation, stomata, anther tapetum development, pollen-stigma recognition, pollen tube guidance and petal shedding and many other important biological processes. According to the current research results, small molecule polypeptide hormones mainly play a role in short-distance signal transduction. They interact with receptor kinases of the LRR-RLK family on the surf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/00C07K7/08
Inventor 刘春明宋秀芬国鹏任仕超徐婷婷
Owner INST OF BOTANY CHINESE ACAD OF SCI
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