摘要
Wnt16属于非经典wnt信号途径上的一员,目前有大量的研究表明wnt16与胚胎的血液发育和骨骼发育相关。为了阐明wnt16在造血过程中的调控机制,利用RT-PCR(逆转录PCR)扩增斑马鱼wnt16基因的ORF(开放阅读框)序列全长,构建真核表达载体pCAGGS-P7/wnt16,测序正确后,用该表达载体免疫BALB/c雌鼠,进行DNA免疫,制备斑马鱼抗WNT16特异性多克隆抗体。结果表明:通过DNA免疫制备的斑马鱼抗WNT16抗血清能特异性识别WNT16蛋白。斑马鱼WNT16抗体成功制备为进一步的功能研究奠定了重要基础。
Wnt16 is a member of the non-classical wnt signaling pathway.Recently,a large number of studies have shown that wnt16 is related to the development of hematopoiesis and skeleton in zebrafish.To elucidate the mechanism of wnt16 in regulation of hematopoiesis,the whole ORF (open reading frame) sequence of wnt16 gene in zebrafish was amplified by RT-PCR (reverse transcription PCR),and the eukaryotic expression vector pCAGGS-P7/ wnt16 was constructed.Then BALB/c female mice immunized with the expression vector to generate anti-WNT16 specific polyclonal antibody in zebrafish.The results showed that the anti-WNT16 antiserum prepared by immunizing DNA could specifically recognize WNT16 protein.The successful preparation of zebrafish WNT16 antibody lays an important foundation for further study.
作者
李玲玉
曹灵慧
欧柏青
江志钢
LI Lingyu;CAO Linghui;OU Baiqing;JIANG Zhigang(The Center for Heart Development,State Key Lab of Development Biology of Freshwater Fish,Hunan Normal University,Changsha 410081,Hunan,China;Geriatric Department,Hunan Provincial People’s Hospital,Changsha 410007,Hunan,China)
出处
《激光生物学报》
CAS
2018年第6期535-539,共5页
Acta Laser Biology Sinica
基金
国家自然科学基金项目(81470449
81700338,81670290)
湖南省自然科学基金项目(06JJ4004)
