title: [Construction and Expression of RNase-Resisting His-Tagged Virus-Like Particles Containing FluA/B mRNA].
abstract: To prepare virus-like particles containing FluA/B mRNA as RNA standard and control in Influenza RNA detection, the genes coding the coat protein and maturase of E. coli bacteriophage MS2 were amplified and cloned into D-pET32a vector. Then we inserted 6 histidines to MS2 coat protein by QuikChange Site-Directed Mutagenesis Kit to construct the universal expressing vector D-pET32a-CP-His. In addition, the partial gene fragments of FluA and FluB were cloned to the down-stream of expressing vector. The recombinant plasmid D-pET32a-CP-His-FluA/B was transformed to BL21 with induction by IPTG. The virus-like particles were purified by Ni+ chromatography. The virus-like particles can be detected by RT-PCR, but not PCR. They can be conserved stably for at least 3 months at both 4 degrees C and -20 degrees C. His-tagged virus-like particles are more stable and easier to purification. It can be used as RNA standard and control in Influenza virus RNA detection.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Xue
authors.forename: Hefei
authors.initials: H
authors.lastname: Xu
authors.forename: Ke
authors.initials: K
authors.lastname: Zhu
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Chen
authors.forename: Juan
authors.initials: J
authors.lastname: Zhang
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lin
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009980:Influenza A virus / Q000235:genetics* / Q000378:metabolism; D009981:Influenza B virus / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012260:Ribonucleases / Q000737:chemistry; D014771:Virion / Q000235:genetics* / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D012260:Ribonucleases