Unlock instant, AI-driven research and patent intelligence for your innovation.

Cordyceps militaris lectin protein Lectin-ccm3, preparation method and application thereof

A technology of lectin and Cordyceps militaris, applied in the field of lectin

Inactive Publication Date: 2017-02-15
SHANGHAI ACAD OF AGRI SCI
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The molecular biological research on lectin in Cordyceps militaris has not been reported yet. The genome of Cordyceps militaris has been sequenced, and it will be annotated as a lectin synthesis gene for prokaryotic expression, and a strain with high expression of lectin will be constructed to clarify the biological activity of lectin in Cordyceps militaris , laying the foundation for the development and utilization of lectins in Cordyceps militaris

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Cordyceps militaris lectin protein Lectin-ccm3, preparation method and application thereof
  • Cordyceps militaris lectin protein Lectin-ccm3, preparation method and application thereof
  • Cordyceps militaris lectin protein Lectin-ccm3, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Sequence Information and Homology Analysis of the Lectin-ccm3 Gene of Cordyceps militaris

[0043] The sequence of the Cordyceps militaris lectin protein Lectin-ccm3 of the present invention is 1299bp, and the detailed sequence is shown in SEQ ID NO.2. The amino acid sequence of Lectin-ccm3 was deduced according to the cDNA sequence, with a total of 432 amino acid residues, a molecular weight of 48.17kDa, a theoretical isoelectric point (pI) of 7.88, an instability coefficient of 32.26, and an aliphatic coefficient of 79.00. See SEQ ID NO for the detailed sequence. 1.

[0044] The amino acid sequence of the Cordyceps brongniartii lectin protein Lectin-ccm3 was searched for protein homology in the Non-redundantGenBank database with the BLAST program, and it was found that the similarity between it and the lectin protein from Cordyceps brongniartii reached 84.79%, The similarity of the lectin protein from Beauveria bassiana was 85.19%, and that of the lectin protein from...

Embodiment 2

[0046] Amplification and expression of Lectin-ccm3 gene from Cordyceps militaris

[0047] The Cordyceps militaris mRNA was used as a template to reverse transcribe into cDNA, and then the cDNA was used as a template to amplify the lectin gene Lectin-ccm3, digested with EcoRI, purified and connected to the pET32a vector digested by EcoRI with ligase (T4ligase) Above, the expression vector pET-Lectin-ccm3 was transformed into the expression strain BL21. Pick a single clone and culture it in LB liquid medium. On the next day, expand the strain at 1:50 to 200 mL, culture at 37°C until OD600=0.4-0.6, add 0.2mM IPTG, and induce expression at 16°C for 12h.

Embodiment 3

[0049] Extraction of Recombinant Cordyceps militaris Lectin-ccm3

[0050] Centrifuge at 8000rpm at 4°C for 5min, collect the induced expression strains, add 1mL of crushing solution to resuspend, freeze with liquid nitrogen and then grind. Centrifuge at 12000rpm, 4°C for 15min, collect the supernatant and precipitate. The expression of the target protein was detected by SDS-PAGE. The target protein Lectin-ccm3 exists in the form of soluble protein, and it is stored at -20°C after being sterilized by a bacterial filter.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
diameteraaaaaaaaaa
control rateaaaaaaaaaa
Login to View More

Abstract

The invention relates to a cordyceps militaris lectin protein Lectin-ccm3, a preparation method and an application thereof. The amino acid sequence is shown as SEQ ID NO.1. The preparation method comprises the following steps: (1) linking cDNA sequence of the cordyceps militaris lectin protein into a vector, thereby obtaining a recombinant vector; (2) transferring the host cells from the recombinant vector, thereby obtaining a recombination strain; (3) culturing the recombination strain and inducing the expression of the recombinant cordyceps militaris lectin protein Lectin-ccm3; (4) purifying, dialyzing and separating, thereby obtaining the Lectin-ccm3. The Lectin-ccm3 is used for preparing an anti-microbial anti-cancer drug. According to the invention, the cordyceps militaris lectin protein Lectin-ccm3 is obtaining through in vitro recombination, the growth of staphylococcus aureus can be restrained, and the biological activity, such as multiplication of human cervical cancer cell HeLa, can be restrained, so that the cordyceps militaris lectin protein Lectin-ccm3 has an application potential at the anti-microbial anti-anticancer aspects.

Description

technical field [0001] The invention belongs to the field of lectins, and in particular relates to a lectin protein Lectin-ccm3 from Cordyceps militaris and its preparation method and application. Background technique [0002] Lectins are a class of non-antibody proteins that can selectively recognize sugars and bind to them non-covalently and reversibly. Because of its ability to agglutinate red blood cells, it is named lectin (Barcmdes, 1988). In 1995, Peumans and Van Damme proposed a new definition of plant lectins, that is, lectin proteins contain at least one non-catalytic domain that can reversibly bind to specific monosaccharides or oligomeric (oligo)saccharides (Peumans, 1995) . [0003] Cell agglutination is the easiest phenomenon to visually characterize the binding of lectins to cells. Lectins generally have two or more sugar binding sites, so they can bind to multiple specific cells at the same time, so that the free cells aggregate into clusters to produce ag...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/37C12N15/31C12N15/70C12N1/21A61K38/16A61P31/04A61P35/00C12R1/19
CPCC07K14/37A61K38/00C12N15/70C12N2800/101
Inventor 鲍大鹏汪滢马元伟高英女王莹李燕唐利华茅文俊周陈力龚明万佳宁杨瑞恒吴莹莹
Owner SHANGHAI ACAD OF AGRI SCI