Expression of vp1 Gene Fragment of Goose parvovirus in Escherichia coli and Preparation of Its Antiserum
WANG Jing;WANG Dong-shuai;HAN Zong-xi;SHAO Yu-hao;RAN Duo-liang;LIU Sheng-wang;KONG Xian-gang
1. National Laboratory of Veterinary Biotechnology, Institute of Harbin eterinary Medicine, Chinese Academy of Agricultural Sciences, Haerbin 150001,China； 2.College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 30052, China; 3. Institute of Gansu Trade Technology ,Lanzhou 730020, China
Abstract：A 662 bp fragment at 5'- end of vp1 gene of Goose parvovirus(GPV) isolate HG5/82 was amplified by PCR. The PCR product was cloned into pMD18-T Simple vector and the recombinant plasmid was transformed into Escherichia coli DH5α. After digesting with BamHⅠand Hind Ⅲ, the inserted fragment was subcloned into prokaryotic expression vector pPROEXTMHTb and the recombinant plasmid was verified by DNA sequencing and transformed into E.coli DH5α. SDS-PAGE results indicated that engineering bacteria could express a 32 kD product after inducing with 0.6 mmol/L IPTG. Densitometric scanning showed the expressed fusion protein could account as much as 22.8% of total bacterial protein of DH5α. The induced engineering bacteria were lysed by 6 mol/L guanidine hydrochloric acid and ultrasound, then the fusion protein was purified with ProBondTM resin from the suspension centrifuged. Antiserum against the fusion protein was obtained by immunizing rabbits. Western blotting analysis showed that the antiserum could specifically recognize the fusion protein as well as VP1 and VP2 of GPV HG5/82 strain.
王静;王懂帅;韩宗玺;卲昱昊;冉多良;刘胜旺;孔宪刚. 鹅细小病毒vp1基因片段的原核表达及抗血清的制备[J]. , 2006, 14(5): 788-792.
WANG Jing;WANG Dong-shuai;HAN Zong-xi;SHAO Yu-hao;RAN Duo-liang;LIU Sheng-wang;KONG Xian-gang. Expression of vp1 Gene Fragment of Goose parvovirus in Escherichia coli and Preparation of Its Antiserum. , 2006, 14(5): 788-792.
