title: Immunolocalization of α18- and α12-giardin in Giardia lamblia trophozoites.
abstract: To study subcellular localization of α18- and α12-giardin in Giardia lamblia trophozoites, the α18- and α12-giardin genes were amplified from G. lamblia assemblage A, respectively. The PCR products were cloned into the prokaryotic expression vector pET-28a(+), and the positive recombinant plasmids were transformed into E. coli Rosetta (DE3) strain for the expression, and expressed α18- and α12-giardin fusion protein were purified by Ni-Agarose resin, respectively. Mice were immunized with purified fusion proteins for preparation of polyclonal antibody, and then the subcellular localization of α18- and α12-giardin was determined by fluorescence immunoassay. Results showed that the concentrations of purified α18- and α12-giardin fusion proteins were 1.20 and 0.86 mg/ml, respectively. The titers of anti-α18- and anti-α12-giardin polyclonal antibody were both as high as 1:25600 dilutions. Immunofluorescent analysis showed that α18- and α12-giardin proteins were mainly localized at four pairs of flagella and the cytoplasm of G. lamblia trophozoites, suggesting that α18- and α12-giardin are the flagella and cytoplasm-associated proteins, respectively. The above information would lay the foundation for research about the crystal structure and biological function of α18- and α12-giardin.
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Wu
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Weida
authors.initials: W
authors.lastname: Pan
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Xianli
authors.initials: X
authors.lastname: Shi
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Auwalu Yusuf
authors.initials: AY
authors.lastname: Abdullahi
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Xingang
authors.initials: X
authors.lastname: Yu
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Biao
authors.initials: B
authors.lastname: Jiang
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Kangxin
authors.initials: K
authors.lastname: Li
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China.
authors.forename: Chang
authors.initials: C
authors.lastname: Xu
authors.affiliation: Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China. gqli@scau.edu.cn.
authors.forename: Guoqing
authors.initials: G
authors.lastname: Li
mesh_terms: D000818:Animals; D003593:Cytoplasm / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D005407:Flagella / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016829:Giardia lamblia / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005873:Giardiasis / Q000469:parasitology*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D051379:Mice; D021381:Protein Transport; D015800:Protozoan Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D053058:Trophozoites / Q000737:chemistry / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C038207:giardin protein, Giardia lamblia
keywords: Giardia lamblia; Prokaryotic expression; Purification; Subcellular localization; α12-giardin; α18-giardin