Chrysalid pteromalid antibiotic protein Pp-AP4 and encoding nucleic acid sequence thereof

An antibacterial protein, nucleic acid sequence technology, applied in the fields of genetic engineering and molecular biology, can solve problems such as reporting antibacterial peptides

Inactive Publication Date: 2008-12-24
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Many researchers have discovered hundreds of antimicrobial proteins from different insects, but t

Method used

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  • Chrysalid pteromalid antibiotic protein Pp-AP4 and encoding nucleic acid sequence thereof
  • Chrysalid pteromalid antibiotic protein Pp-AP4 and encoding nucleic acid sequence thereof
  • Chrysalid pteromalid antibiotic protein Pp-AP4 and encoding nucleic acid sequence thereof

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Experimental program
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Embodiment 1

[0092] Embodiment 1: (with reference to Fig. 1:)

[0093] 1. Construction of Chrysalis chrysalis cDNA (complementary DNA) expression library:

[0094] Chrysalis chrysalis came from the Laboratory of Insect Physiology and Biochemistry, Institute of Insect Science, Zhejiang University.

[0095] a. Isolate total RNA from the whole body of Chrysalis chrysalis, obtain poly(A)+RNA with purified mRNA kit, and then use cDNA synthesis kit, Synthesis kits and A cDNA (complementary DNA) expression library was constructed using the Gigapack III cloning kit (purchased from the Takana agency of American clontech company, without a Chinese name).

[0096] b. According to the operation instructions of the kit, each step is performed to construct a cDNA (complementary DNA) expression library.

[0097] 2. Screening of Chrysalis chrysalis cDNA (complementary DNA) expression library:

[0098] a. Prepare SOLR (a type of Escherichia coli, no Chinese name) bacterial solution: burn the inoculat...

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Abstract

The invention discloses a nucleic acid sequence of a pteromalus puparum antifungal protein Pp-AP4 code. The separated DNA molecule comprises: a polypeptide nucleotide sequence with the pteromalus puparum antifungal protein Pp-AP5 activity is encoded, the nucleotide sequence and the nucleotide sequence from the 226th ribonucleotide to the 345th ribonucleotide in the SEQ ID NO:1 have 70 percent of homology; or the nucleotide sequence can be hybridized with the nucleotide sequence from the 226th ribonucleotide to the 345th ribonucleotide in the SEQ ID NO:1 at a temperature of between 40 and 55 DEG C. The invention also discloses a pteromalus puparum antifungal protein polypeptide Pp-AP4 which has an amino acid sequence shown in the SEQ ID NO: 2. The pteromalus puparum antifungal protein Pp-AP4 has the antibiosis performance.

Description

technical field [0001] The invention relates to the fields of molecular biology and genetic engineering. In particular, it is an antibacterial protein Pp-AP4 expressed in the chrysalis chrysalis and its coded nucleic acid sequence. Background technique [0002] In recent years, due to the increasing problems of drug abuse, drug residues and bacterial resistance, existing antibiotics are losing their original efficacy, which has aroused people's attention to food safety, and more and more countries have begun to call for banning Antibiotics, and antibacterial peptides (antibacterial peptides) have aroused great research interest because of their unique biological activity and special mechanism of action different from traditional antibiotics, and have become one of the hot spots in the field of molecular biology and biochemistry research. Researchers from various countries have discovered hundreds of antimicrobial peptides from mammals, insects, amphibians, etc., some of whi...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/435A23K1/16A61K38/17A61P31/00
Inventor 慎小晶叶恭银成雄鹰胡萃
Owner ZHEJIANG UNIV
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