LANA viral protein binding sites
a technology of lana virus and binding site, which is applied in the field of lana virus protein binding site, can solve problems such as interfering with host processes, and achieve the effects of preventing transcription, strengthening interaction, and closer complementarity to perfection
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Identification of LANA binding Sequences in Viral Genomes
[0158] A. Identification of Potential Consensus LANA-Binding Sites via Randomer Immunoprecipitation
[0159] In an effort to identify potential LANA-binding sites, a template for PCR amplification was synthesized which included KpnI and EcoRI restriction sites flanking a random 40-bp oligomer. Sense and antisense primers, which annealed to the nonrandom KpnI / EcoRI flanking sequences, were used to amplify a large pool of random 40-bp oligomers. This DNA was purified and mixed with anti-myc immunoprecipitates from 293 cells stably transduced with a myc-tagged LANA expression construct. After a period of 30 min, the immunoprecipitates were centrifuged out of the binding reaction and washed four times in binding buffer. These beads were then resuspended, heated at 95.degree. C. for 10 min, and then extracted with phenol:chloroform followed by chloroform isoamyl alcohol. Oligomers that had been precipitated with the LANA protein were ...
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