血吸虫极低密度脂连结蛋白的表达及免疫原性鉴定
摘要
目的：在大肠杆菌中表达日本血吸虫特异性极低密度脂连结蛋白(SVLBP)，纯化并鉴定其免疫活性。方法：SVLBP基因克隆菌在IPTG诱导下大肠杆菌中以6个组氨酸融合蛋白形式大量表达；非变性条件下制备克隆菌裂解液，离心后上演液经凝胶亲和层析纯化重组蛋白，已纯化的重组SSVLBP免疫家兔制备特异性抗血清，用SDS—PAGE和Western blot鉴定其免疫活性。结果：克隆菌在IPIG诱导下能大量表达重组蛋白SVLBP；重组SSVLBP蛋白能诱导家兔产生单特异抗体，琼脂双扩散法测抗体滴度＞1：16；表达产物能与血吸虫病人血清反应；单特异抗血清能识别天然成虫可溶性膜抗原的单一蛋白。结论：SVLBP能在克隆菌中大量表达并且具有较好的免疫原性。
参考文献10
-
1Meyer F, Meyer H, Bueding E.Lipid in the parasitic and free- living flatworms. Biochem Biophys Acta, 1970,210:257-266.
-
2Rogers MV, Davem KM, Smythe JA, et al.Immunoblotting analysis of the major integral membrane protein antigens of Schistosoma japonicum.Mol Biochem Parasitol , 1988, 29:77-78.
-
3Pogers MV, Quilici D,Mitchell GF, et al.Purification of a putative Lipoprotein receptor from Schistosoma japonicum adult worms .Mol Biochem Parasital, 1990, 41:93-100.
-
4Chiang CP, Caulifield JP.The binding of human low - density lipoproteins to surface of schistosomula of Schistosoma mansoni is inhibited by polyanions and reduces the binding of anti-schistosomal antibodies .Am J Pathol, 1989, 134(5):1007-- 1018.
-
5Chiang OP, Caulifield JP.Human lipoprotein binding to schistosomula of Schistosoma mansoni .Displacement by polyanions , parasite antigen masking, and pezsistence in young larvae. Am J Pathol, 1989,135(6):1015-1024.
-
6Bennett MW, Caulfield.Specific binding of human low-density lipoprotein to the surface of schistosomula of Schistosoma mansoni and ingestion by the parasite.Am J Pathol, 1991,138(5) : 1173-1182.
-
7Sambrook J, Fritsch EF, Maniatis T, et al. Molecular Cloning, a Laboratory Nanual.2nd ed.New York: Cold sprng Harbor.Laboratory. Cold Spring Harbor, 1989, 880-- 898.
-
8Ruppel A, Disfeld HJ, Roth er U.Immunoblot analysis of Schistosoma mansoni antigens with sera of schistosomiasis patients:diagnostic potential of an adult schistosome polypeptide. Chin Exp Immunol, 1985, 62: 499- 506.
-
9Hancock, Mohamed YB, Xue HC, et al.A recombinant protein from Schistoma maasoni useful for the detection of S.mansoni and Schistoma haematobium antibodies. J Parasitol, 1997, 83:612-618.
-
10Liu SX, Song GC, Xu YX, et al.Immunization of mice with recombinnat Sjc26GST induces a pronouced anti - fecundity effect after experimental infection with Chinese Schistosoma japonicum. Vaccine, 1995,13(6) : 603-- 607.
-
1干小仙,王越,曾肖芃,丁建祖,沈慧英,沈丽英,PaulBrindley,樊晋江.重组日本血吸虫极低密度脂结合蛋白的抗原性及诊断价值的研究[J].浙江省医学科学院学报,2005,16(2):30-33.
-
2王延风,冯宗忱.兔主动脉平滑肌细胞代谢正常极低密度脂蛋白的机制[J].宜昌医学专科学校学报,1990,18(2):105-108.
-
3王延风,王式平.巨噬细胞促进兔主动脉平滑肌细胞摄取极低密度...[J].生物化学与生物物理学报,1992,24(1):33-39.
-
4申鸣,钟显春,颜炜群,宇莉,侯立中,杨同书.软骨型PG核心蛋白透明质酸结合区和连结蛋白的提纯及鉴定[J].中国地方病防治,1997,12(4):219-221.
-
5王恩芳,刘曙玮.番茄红素通过激活PI3K/Akt信号通路对2型糖尿病小鼠治疗机制研究[J].实用糖尿病杂志,2015,11(5):35-37. 被引量：5
-
6张桂筠,张兆松,沈一平,陈淑贞,吴海玮,苏川,吴观陵.日本血吸虫重组抗原的免疫原性鉴定[J].中国人兽共患病杂志,1997,13(5):28-30. 被引量：2
-
7张志祥.血脂高的形成及预防[J].中国机关后勤,2001,0(11):39-39.
-
8王越,干小仙,曾肖芃,沈慧英,丁建祖,沈丽英,樊晋江.SVLBP重组抗原用于日本血吸虫病免疫诊断的研究[J].热带医学杂志,2004,4(5):518-520. 被引量：9
-
9李巍,冯宗忱,邓耀祖.巨噬细胞对正常人极低密度脂蛋白亚组分代谢的研究[J].生物化学杂志,1993,9(3):287-292.
-
10陈培利,冯友梅,从容,王淳本,宗义强,邓耀祖.极低密度脂蛋白对小鼠腹腔巨噬细胞载脂蛋白E基因表达的影响[J].中国动脉硬化杂志,1999,7(2):95-98. 被引量：3
