Zhou Ji-Yong, Cheng Li-Qin, Zheng Xiao-Juan, Wu Jian-Xiang, Shang Shao-Bin, Wang Jin-Yong, Chen Ji-Gang
Institute of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310029, China.
J Biotechnol. 2004 Jul 15;111(2):121-30. doi: 10.1016/j.jbiotec.2004.03.012.
To seek a new delivery system of vaccine for infectious bronchitis virus (IBV), transgenic potato expressing full-length spike (S) protein of IBV was produced and its immunogenicity in chickens was investigated. One to three copies of S gene of IBV were randomly and stably inserted into potato (Solanum tuberrosum cv. Dongnong 303) genome by Agrobacterium tumefaciens-mediated transformation. Transcription and translation of S gene for IBV were confirmed by Northern blot and Western blot analyses in transgenic plantlets. Chickens immunized orally and intramuscularly with transgenic potato tubers expressing S protein generated the detectable levels of serum neutralizing antibodies and were protected against the challenge with the virulent IBV. In vitro secretion of interleukin 2 and T lymphocyte proliferation of spleen cells from the immunized chickens varied with the dose and the manner of vaccination with S protein derived from transgenic plants. The results indicated that S protein expressed in transgenic plants might be a new source for the production of Coronaviridae IBV vaccine.
为寻找传染性支气管炎病毒（IBV）疫苗的新型递送系统，制备了表达IBV全长刺突（S）蛋白的转基因马铃薯，并研究了其在鸡体内的免疫原性。通过根癌农杆菌介导的转化，将1至3个拷贝的IBV S基因随机且稳定地插入马铃薯（Solanum tuberrosum cv. Dongnong 303）基因组中。通过Northern印迹和Western印迹分析在转基因小植株中证实了IBV S基因的转录和翻译。用表达S蛋白的转基因马铃薯块茎经口服和肌肉注射免疫的鸡产生了可检测水平的血清中和抗体，并受到了强毒IBV攻击的保护。免疫鸡脾脏细胞的白细胞介素2体外分泌和T淋巴细胞增殖随转基因植物来源S蛋白的接种剂量和方式而变化。结果表明，转基因植物中表达的S蛋白可能是冠状病毒科IBV疫苗生产的新来源。
