title: Sequence analysis and prokaryotic expression of Giardia lamblia α-18 giardin gene.
abstract: To study the genetic variation and prokaryotic expression of α18 giardin gene of Giardia lamblia zoonotic assemblage A and host-specific assemblage F, the α18 genes were amplified from G. lamblia assemblages A and F by PCR and sequenced. The PCR product was cloned into the prokaryotic expression vector pET-28a(+) and the positive recombinant plasmid was transformed into Escherichia coli Rosetta (DE3) strain for the expression. The expressed α18 giardin fusion protein was validated by SDS-PAGE and Western blot analysis, and purified by Ni-Agarose resin. The putative sequence of α18 giardin amino acid was analyzed by bioinformatics software. Results showed that the α18 giardin gene was 861 bp in length, encoding 286 amino acids; it was 100% homologous between human-derived and dog-derived G. lamblia assemblage A, but it was 86.8% homologous with G. lamblia assemblage F (cat-derived). Giardin α18 was about 36 kDa in molecular weight, with good reactivity. Prediction based on in silico analyses: it had hydrophobicity, without signal peptide and transmembrane domain, and contained 11 alpha regions, 13 beta sheets, 1 beta turn and 7 random coils in secondary structure. The above information would lay the foundation for research about the subcellular localization and biological function of α18 giardin in G. lamblia.
authors.affiliation: 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: 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: 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: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province 510642, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
authors.affiliation: 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: 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: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province 510642, People's Republic of China.
authors.forename: Liping
authors.initials: L
authors.lastname: Tan
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province 510642, People's Republic of China.
authors.forename: Meiran
authors.initials: M
authors.lastname: Song
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province 510642, People's Republic of China. Electronic address: gqli@scau.edu.cn.
authors.forename: Guoqing
authors.initials: G
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D019295:Computational Biology; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D016829:Giardia lamblia / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D003598:Cytoskeletal Proteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C038207:giardin protein, Giardia lamblia
keywords: Bioinformatics analysis; Giardia lamblia; Prokaryotic expression; Purification; α18 giardin