Contraction of rat TRESK gene recombinant adenovirus vector

VernacularTitle:大鼠TRESK基因重组腺病毒载体的构建
Author: Jun ZHOU ; Shanglong YAO ; Chengxiang YANG ; Jiying ZHONG ; Hanbing WANG ; Wenjing LIN ; Runxing GAO
Publication Type:Journal Article
Keywords: Potassium channels,tandem pore domain; Adenoviridae; Genes
From: Chinese Journal of Anesthesiology 2011;31(3):296-298
CountryChina
Language:Chinese
Abstract: Objective To construct rat TRESK gene recombinant adenovirus expression vector.Methods TRESK full-length cDNA was cloned from rat dorsal root ganglion cells and confirmed by RT-PCR and sequencing. pAd/CMV/V5-DEST-TRESK was then constructed under the control of CMV-promotor. DH5α colibacillus was translated and the positive recombinants were subsequently identified by PCR and DNA sequencing. 293T cells were cotransfected and packed to produce adenovirus. Results The titer of virus was tested using Hole-by-dilution titer method. The full length of TRESK from rat dorsal root ganglion cells is 781 bp. It was demonstrated that the DNA sequencing was completely consistent with TRESK sequencing of rat recorded in GeneBank. The PCR amplification of the pAd/CMV/V5-DEST-TRESK gDNA was matched with pAD-GFP blank vector as anticipated. The titer of the concentrated virus was 1.31×109 TU/ml. Conclusion Rat TRESK gene recombinant adenovirus vector is constructed successfully.