Coding sequence of arisaema agglutinant protein and its application

A kind of Tiannanxing, coding technology, applied in the field of preparation of the protein and nucleic acid sequence

Inactive Publication Date: 2006-06-14
FUDANDIEN BIOTECH CO LTD SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Before the publication of the present invention, there has not been any publicity or report of the Araceae ah-Lectin protein sequence and its nucleic acid sequence mentioned in this patent application

Method used

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  • Coding sequence of arisaema agglutinant protein and its application
  • Coding sequence of arisaema agglutinant protein and its application
  • Coding sequence of arisaema agglutinant protein and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Cloning of ah-Lectin gene from Araceae

[0056] 1. Tissue separation (isolation)

[0057] The stems of Araceae come from Shanghai University of Traditional Chinese Medicine. After taking the inflorescence material of Aracea inflorescences, they are immediately frozen in liquid nitrogen. .

[0058] 2. RNA isolation (RNA isolation)

[0059] Take part of the tissue, grind it with a mortar, add it to a 50mL tube filled with lysate, shake it fully, and then transfer it into a glass homogenizer. After homogenization, transfer to a new 50ml tube and extract total RNA (TRIzol Reagents, Gibco, NY, USA). The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis.

[0060] 3. Cloning of Full-length cDNA

[0061] According to the conserved amino acid sequences of snowdrops and other araceae lectins, degenerate primers were designed, and the principle of homologous gene cloning was used to adopt the RACE method (GibcoBRL kit)

[0062] Perform cDNA f...

Embodiment 2

[0072] Sequence information and homology analysis of ah-Lectin gene in Araceae

[0073] The length of the novel Aracea ah-Lectin full-length cDNA of the present invention is 1169 bp, and the detailed sequence is shown in SEQ ID NO.3, wherein the open reading frame is located at nucleotides 85-861. The amino acid sequence of Aracea ah-Lectin was deduced according to the full-length cDNA, which has a total of 258 amino acid residues, a molecular weight of 28464 Daltons, and a pI of 7.71. See SEQ ID NO.4 for the detailed sequence.

[0074] The full-length cDNA sequence of ah-Lectin and its encoded protein were identified by BLAST program in the Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR databases. Source search found that it has a certain homology with the gene of Pinellia lectin (PTA) of Araceae (Yao et al., Acta Fudan Sinica 2001, 40: 461-464) and the gene of GNA of Amaryllidaceae sex. At the nucleotide level, i...

Embodiment 3

[0076] Study on the Structure and Function of Araceae ah-Lectin Protein

[0077] 1. Amino acid sequence domain analysis of Aracea ah-Lectin protein. The amino acid sequence of Aracea ah-Lectin protein was searched for domains in the NCBI database (web address: http: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi), and the following results were obtained:

[0078]

[0079] (1) In the amino acid sequence, the following domain regions are present:

[0080] Boxed region (27-131, 148-252): Lectin protein functional module (Lectin).

[0081] (2) Function analysis

[0082] There is a functional module of Lectin protein in the amino acid sequence of the gene, so it can be predicted that it does have the corresponding function.

[0083] 2. Signal peptide analysis of araceae lectin protein. Through the comparative analysis of the nucleotide sequence and amino acid sequence of the araceae lectin protein and other plant lectins, it was found that the gene of the araceae lectin prote...

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Abstract

The invention provides a new ah-Lectin protein expressed in Araceae, its coding sequence and its application in improving plant resistance to diseases and insect pests. The present invention relates to a fusion gene construct comprising said gene, and a new recombinant expression vector carrying the construct. It also relates to plant cells transformed with said expression vector, and said gene transgenic plants produced by transformed cells and their progeny, including plant seeds and plant tissues. The invention expresses the gene in plants, and the obtained transgenic plants have enhanced resistance to plant diseases and insect pests.

Description

technical field [0001] The invention relates to the fields of molecular biology, enzymology, physiology, genetic engineering and the like. Specifically, the present invention relates to an ah-Lectin protein (Arisaemaheterophyllum agglutinin, AHA) expressed in Araceae and its nucleic acid sequence. The present invention also relates to the preparation method and use of the protein and nucleic acid sequence. Background technique [0002] In the past 20 years, the research on the application of lectins in biology has developed rapidly. Its biggest feature is that it can recognize sugars and sugars. Each lectin has the ability to specifically bind to a special carbohydrate (Ann Biochem, 1981, 50: 207-231). Certain lectin genes have strong anti-Homoptera insects including aphids and planthoppers (Entomol Exp Appl, 1993, 66: 119-126; Transgenic Res, 1995, 4: 18-25; Entomol Exp Appl, 1995, 75:51-59). [0003] In the present invention, we cloned ah-Lectin, a homologous gene of t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/29C12N15/63C12N15/64C07K14/12C07K16/16C07H21/00C12Q1/68A01H1/00
Inventor 唐克轩赵秀云孙小芬廖志华游学军
Owner FUDANDIEN BIOTECH CO LTD SHANGHAI
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