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甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) 血凝素 (Hemagglutinin / HA) (Codon Optimized) ORF mammalian expression plasmid, Flag 标签

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H6N1 HA cDNA Clone产品信息
NCBI注册码:
参考序列ORF长度:1701bp
cDNA基因描述:Full length Clone DNA of Influenza A H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin DNA.
分子别称:HA
分子种属:H6N1
载体:pCMV/hygro-FLAG
质粒:pCMV-H6N1-NC-07-HA-flag
限制性酶切位点:KpnI + XhoI (5.5kb + 1.73kb)
Tag序列:FLAG Tag Sequence: GATTACAAGGATGACGACGATAAG
序列信息:A number of silent mutations were introduced into the DNA sequence in order to increase its protein expression level in mammalian cell system. The translated amino acid sequence is identical with ACE81692.1.
测序引物:T7(TAATACGACTCACTATAGGG) BGH(TAGAAGGCACAGTCGAGG)
启动子:Enhanced CMV mammalian cell promoter
Application:Stable or Transient mammalian expression
抗生素(大肠杆菌):Kanamycin
抗生素(哺乳动物细胞):Hygromycin
运输方式:Each tube contains lyophilized plasmid.
储存方法:The lyophilized plasmid can be stored at ambient temperature for three months.
pCMV/hygro-FLAG Vector Information
 
Vector Name pCMV/hygro-FLAG
Vector Size 5681bp
Vector Type Mammalian Expression Vector
Expression Method Constiutive ,Stable / Transient
Promoter CMV
Antibiotic Resistance Ampicillin
Selection In Mammalian Cells Hygromycin
Protein Tag FLAG
Sequencing Primer Forward:T7(TAATACGACTCACTATAGGG)
Reverse:BGH(TAGAAGGCACAGTCGAGG)

Schematic of pCMV/hygro-FLAG Multiple Cloning Sites

FLAG Tag Info

FLAG-tag, or FLAG octapeptide, is a polypeptide protein tag that can be added to a protein using recombinant DNA technology. It can be used for affinity chromatography, then used to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. It can also be used in the isolation of protein complexes with multiple subunits.

A FLAG-tag can be used in many different assays that require recognition by an antibody. If there is no antibody against the studied protein, adding a FLAG-tag to this protein allows one to follow the protein with an antibody against the FLAG sequence. Examples are cellular localization studies by immunofluorescence or detection by SDS PAGE protein electrophoresis.

The peptide sequence of the FLAG-tag from the N-terminus to the C-terminus is: DYKDDDDK (1012 Da). It can be used in conjunction with other affinity tags, for example a polyhistidine tag (His-tag), HA-tag or myc-tag. It can be fused to the C-terminus or the N-terminus of a protein. Some commercially available antibodies (e.g., M1/4E11) recognize the epitope only when it is present at the N-terminus. However, other available antibodies (e.g., M2) are position-insensitive.

甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin ORF mammalian expression plasmid (Codon Optimized), C-GFPSpark-taggedVG11723-ACG2070
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin (Codon Optimized) ORF mammalian expression plasmid, C-OFPSpark / RFP 标签VG11723-ACR2070
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin ORF mammalian expression plasmid (Codon Optimized)VG11723-C1890
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin ORF mammalian expression plasmid (Codon Optimized), C-Flag 标签VG11723-CF1890
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin (Codon Optimized) ORF mammalian expression plasmid, C-His 标签VG11723-CH1890
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin (Codon Optimized) ORF mammalian expression plasmid, C-Myc 标签VG11723-CM1890
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin (Codon Optimized) ORF mammalian expression plasmid, C-HA 标签VG11723-CY1890
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin (Codon Optimized) ORF mammalian expression plasmid, N-Flag 标签VG11723-NF1890
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin (Codon Optimized) ORF mammalian expression plasmid, N-His 标签VG11723-NH1890
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin ORF mammalian expression plasmid (Codon Optimized), N-Myc-taggedVG11723-NM1890
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin (Codon Optimized) ORF mammalian expression plasmid, N-HA 标签VG11723-NY1890
甲型流感 H6N1 (A/northern shoveler/California/HKWF115/07) Hemagglutinin ORF mammalian expression plasmid (Codon Optimized)VG11723-UT1890
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研究背景

The influenza viral Hemagglutinin (HA) protein is a homo trimer with a receptor binding pocket on the globular head of each monomer.HA has at least 18 different antigens. These subtypes are named H1 through H18.HA has two functions. Firstly, it allows the recognition of target vertebrate cells, accomplished through the binding to these cells' sialic acid-containing receptors. Secondly, once bound it facilitates the entry of the viral genome into the target cells by causing the fusion of host endosomal membrane with the viral membrane.The influenza virus Hemagglutinin (HA) protein is translated in cells as a single protein, HA0, or hemagglutinin precursor protein. For viral activation, hemagglutinin precursor protein (HA0) must be cleaved by a trypsin-like serine endoprotease at a specific site, normally coded for by a single basic amino acid (usually arginine) between the HA1 and HA2 domains of the protein. After cleavage, the two disulfide-bonded protein domains produce the mature form of the protein subunits as a prerequisite for the conformational change necessary for fusion and hence viral infectivity.

参考资料
  • White JM, Hoffman LR, Arevalo JH, et al. (1997). "Attachment and entry of influenza virus into host cells. Pivotal roles of hemagglutinin". In Chiu W, Burnett RM, Garcea RL. Structural Biology of Viruses.
  • Suzuki Y (March 2005). "Sialobiology of influenza: molecular mechanism of host range variation of influenza viruses". Biol. Pharm. Bull. 28 (3): 399–408.
  • Senne DA, Panigrahy B, Kawaoka Y, et al. (1996). "Survey of the hemagglutinin (HA) cleavage site sequence of H5 and H7 avian influenza viruses: amino acid sequence at the HA cleavage site as a marker of pathogenicity potential". Avian Dis. 40 (2): 425–37
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