鸡毒支原体licA基因编码脂寡糖/脂多糖(lipooligosaccharide/lipopolysaccharide,LOS/LPS)合酶或胆碱激酶,催化合成磷酸胆碱。该产物是支原体表面脂质的主要组成部分,与其致病性密切相关。鸡毒支原体的licA基因位于hmw3-licA-mgc2-gapA-crmA-crmB-crmC粘附素基因簇内部。为深入研究licA基因及其产物的生物学活性,本研究对鸡毒支原体licA基因进行了真核表达,通过间接免疫荧光试验检测,结果发现LicA主要定位于杆状病毒和DF1细胞浆中,并不对sf9和DF1真核细胞具有明显的毒性。这为下一步在体外建立LicA生物学活性检测的模型奠定了基础。
The licA gene of Mycoplasma gallisepticum encodes lipooligosaccharide / lipopolysaccharide synthase,or choline kinase catalyzing choline to phosphorylcholine,which is the major component of membrane lipids and associated with the pathogenicity of mycoplasmas.The licA gene is located in the adhesion gene cluster(hmw3-licA-mgc2-gapA-crmA-crmB-crmC).In this study,the licA gene was amplified and subcloned into Bac-to-Bac Baculovirus eukaryotic expression system and pcDNA3.1 vector.The LicA protein was expressed and sublocalized in the cytoplasma of the transfected sf9 and DF1 cells without cytotoxicity as demonstrated in Western blot analysis and indirect immunofluoscence assay.