Arisaema amurense maxim agglutinin protein coding sequence and application thereof
A kind of Tiannanxing, coding technology, applied in the field of lectin protein and its nucleic acid sequence
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0038] Cloning of ArA-Lectin Gene from Northeast Araceae
[0039] 1. Tissue Isolation
[0040] The Northeast Aracea tuber (a common plant) was collected from the Changbai Mountain area in Jilin Province, China. After the tuber was harvested, it was immediately frozen and stored in general-purpose liquid nitrogen.
[0041] 2. Extraction of RNA
[0042] The RNA extraction method refers to the instruction of TRIzol (see: TRIzol Reagents, Gibco, NY, USA). The specific steps are: 1) Weigh 0.5 gram of Northeast Ariana tuber, grind it into fine powder with liquid nitrogen, and distribute it in two 1.5ml Eppendorf tubes (as fast as possible to avoid RNA degradation);
[0043] 2) Add 1ml TRIzol to each tube and shake vigorously to mix it evenly, and place it at room temperature for 5min (the amount of the sample should not exceed 10% of TRIzol);
[0044] 3) Centrifuge at 12000rpm for 10min at 4°C, and pipette the supernatant into a clean 1.5ml Eppendorf tube;
[0045] 4) Add 0.2ml ...
Embodiment 2
[0120] Sequence Information and Homology Analysis of the ArA-Lectin Gene of Northeast Araceae
[0121] The length of the open reading frame of the novel ArA-Lectin of the present invention is 780bp, and the detailed sequence is shown in SEQ ID NO:1. The amino acid sequence of ArA-Lectin was deduced according to the full-length cDNA. It has a total of 259 amino acid residues, a molecular weight of 28.303 Daltons, and an isoelectric point pI of 7.07. Its amino acid sequence is shown in SEQ ID NO:1.
[0122] The full-length cDNA sequence of ArA-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 showed that it has high homology with Araceae plant lectins (Yao et al., 2001). At the amino acid level, it has 85% homology with Pinellia trifoliate agglutinin (PTA) protein, 82% homology with Pinellia palmata agglutinin (PPA) protein, a...
Embodiment 3
[0124] Amino acid sequence domain analysis of ArA-Lectin protein from Northeast Araceae.
[0125] The amino acid sequence of the ArA-Lectin protein in Northeast Araceae was searched for the structural domain in the NCBI database (website: http / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi), and it was found that in the ArA-Lectin protein There are two conserved domains (results such as image 3 ), both are mannose-binding domains. Domain A is located at T 27 and R 138 Between amino acids, domain B is located at D 148 and G 252between amino acids. And the two structural domains have high similarity. The high-level structure of ArA-Lectin protein was analyzed with VectorNTI Suite 6.0. ArA-Lectin protein consists of 12 β-sheets, and the β-sheets are connected by turns and coils ( Figure 4 ). The signal peptide forms an alpha helix. ArA-Lectin protein overall forms a very strong structure, which is highly similar to the three-dimensional structure of other mannose-bindi...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More