發(fā)布時間：2018-05-31 10:31

  本文選題：反義寡核苷酸 + 大腸埃希菌��； 參考：《中國獸醫(yī)學(xué)報》2017年02期

【摘要】：為探討反義寡核苷酸(ASODN)沉默acrA、marA和acrD基因前后,黃芩苷對臨床耐藥大腸埃希菌(E.coli)環(huán)丙沙星敏感性的影響,設(shè)計并合成3個基因的ASODN引物,設(shè)置空白對照組、黃芩苷組、ASODN沉默組、ASODN沉默+黃芩苷組,分別轉(zhuǎn)染到臨床耐藥菌感受態(tài)細(xì)胞中,涂布于一定濃度環(huán)丙沙星平板上,培養(yǎng)觀察菌落生長情況并計數(shù)。結(jié)果發(fā)現(xiàn),相對于空白對照組,黃芩苷組菌落數(shù)顯著降低;acrA、acrD和marA基因沉默組與空白對照組相比,除了acrD基因沉默1號菌株和marA基因沉默的Y17菌株外,其余菌株在環(huán)丙沙星平板上的菌落數(shù)也出現(xiàn)不同程度地下降,具有顯著性差異;acrA、acrD和marA基因沉默+黃芩苷組與黃芩苷組相比,多數(shù)菌株在環(huán)丙沙星平板上的菌落數(shù)均出現(xiàn)不同程度地下降;acrA基因沉默+黃芩苷組與acrA基因沉默組相比,4株菌株變化差異不顯著,Y17號菌株菌落數(shù)有所上升,J45號菌株在該基因沉默后,環(huán)丙沙星平板上的菌落數(shù)出現(xiàn)下降的趨勢,差異顯著;acrD基因沉默+黃芩苷組與acrD基因沉默組相比,除了1號菌株變化差異不顯著外,其余5株菌株變化差異顯著,但Y17號菌株菌落數(shù)上升;marA基因沉默+黃芩苷組與marA基因沉默組相比,除了Y17、Y35這2株菌株變化差異顯著外,其余4株菌株變化差異不顯著。以上結(jié)果表明,黃芩苷主要通過抑制acrA和marA發(fā)揮外排抑制作用,acrD不是其增強(qiáng)環(huán)丙沙星敏感性的主要靶點(diǎn)。
[Abstract]:In order to investigate the effect of baicalin on the susceptibility of clinical resistant Escherichia coli to ciprofloxacin before and after antisense oligonucleotide silencing acrAlomara and acrD genes, ASODN primers of three genes were designed and synthesized, and blank control group was set up. Baicalin group was silenced by ASODN silencing baicalin group and transfected into the competent cells of clinical drug-resistant bacteria and coated on a certain concentration of ciprofloxacin plate to observe the colony growth and count. The results showed that the colony number of baicalin group was significantly lower than that of blank control group, except acrD gene silencing strain 1 and marA gene silencing strain Y17. The colony number of other strains on ciprofloxacin plate also decreased to some extent, and there was significant difference between baicalin group and baicalin group in acrAgnacrD and marA gene silencing. The colony number of most strains on ciprofloxacin plate was decreased in varying degrees. There was no significant difference between acrA gene silencing baicalin group and acrA gene silencing group. The colony number of Y17 strain increased to J45. After the gene was silenced, The number of colonies on ciprofloxacin plate decreased. The difference was significant between baicalin silencing group and acrD gene silencing group, except for strain 1, there were significant differences among the other 5 strains. However, there was no significant difference between Y17 and marA gene silencing baicalin group, except Y17 Y35 strain, except Y17 Y35 gene silencing baicalin group and Y17 Y35 gene silencing group, but there was no significant difference in the other 4 strains in comparison with the marA gene silencing group. These results indicate that baicalin exerts efflux inhibition mainly by inhibiting acrA and marA. AcrD is not the main target for enhancing ciprofloxacin sensitivity.
【作者單位】： 河南農(nóng)業(yè)大學(xué)牧醫(yī)工程學(xué)院;
【基金】：國家自然科學(xué)基金資助項(xiàng)目(U1304328) 河南省科技廳資助項(xiàng)目(132300410113)
【分類號】：R285
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本文編號：1959306