Cloning and Sequence Analysis of TpRS1 Gene from Cysticercus pisiformis

Abstract

Abstract: To study the function of TpRS1 gene from Cysticercus pisiformis, the open reading frame（ORF）cDNA sequence of TpRS1 gene was cloned by RT-PCR from C. pisiformis total RNA with primers derived from Taenia solium RS1 gene sequence in the GenBank database. Biochemical properties, secondary structure, signal, hydrophobicity and antigenicity in TpRS1 protein were predicted by bioinformatics tools. The results indicated that TpRS1 cDNA sequence contained an ORF of 258 nucleotides and the deduced protein consisted of 85 amino acids with the theoretical molecular weight of 9.6 kD and isoelectric point of 9.07. Analysis of secondary structure revealed 70.59% and 5.88% of α-helix and β-strands, respectively, and others were loop. The signal peptide sequence of TpRS1 were not identified. TpRS1 protein sequence showed more than 72% identity with other Taenia spp.. Phylogenetic analysis indicated that TpRS1 located in the same clade with Taenia hydatigena.

Key words: Cysticercus pisiformis, TpRS1 gene, Cloning, Sequence analysis

Han Lele, Li Zuolei, Gou Huitian, Sun Xiaolin. Cloning and Sequence Analysis of TpRS1 Gene from Cysticercus pisiformis[J]. Biotechnology Bulletin, 2014, 0(7): 131-136.

References

[1] 汪明.家畜寄生虫学［M] . 第3版.北京：中国农业出版社, 2003：117-118.
[2] Toral-Bastida E, Garza-Rodriguez A, Jimenez-Gonzalez DE, et al. Development of Taenia pisiformis in golden hamster（Mesocricetus auratus）[J]. Parasit Vectors, 2011, 4：147.
[3] Kyngdon CT, Gauci CG, Rolfe RA, et al. In vitro oncosphere-killing assays to determine immunity to the larvae of Taenia pisiformis, Taenia ovis, Taenia saginata, and Taenia solium[J]. J Parasitol, 2006, 92（2）：273-281.
[4] 杨世广, 汪志楷, 施宝坤.豆状带绦虫的人工感染及其发育史[J].中国兽医科技, 1986, 16（6）：16-19.
[5] 周永学, 杜爱芳, 张雪娟, 等.我国兔豆状囊尾蚴病的研究进展[J].中国养兔杂志, 2006（1）：25-28.
[6] 张守发, 鞠玉琳, 李学一, 等.不同抗原在兔豆状囊尾蚴病血清学诊断中的研究[J].延边大学农学学报, 1994, 16（2）：98-102, 108.
[7] Ferrer E, Bonay P, Cuevas MF, et al. Molecular cloning and characterisation of Ts8B1, Ts8B2 and Ts8B3, three new members of the Taenia solium metacestode 8 ku diagnostic antigen family[J]. Mol Biochem Parasitol, 2007, 152（1）：90-100.
[8] Assana E, Kanobana K, Tume CB, et al. Isolation of a 14 ku antigen from Taenia solium cyst fluid by HPLC and its evaluation in enzyme linked immunosorbent assay for diagnosis of porcine cysticercosis[J]. Res Vet Sci, 2007, 82（3）：370-376.
[9] Hancock K, Khan A, Williams FB, et al. Characterization of the 8-kilodalton antigens of Taenia solium metacestodes and evaluation of their use in an enzyme-linked immunosorbent assay for serodiagnosis[J]. J Clin Microbiol, 2003, 41（6）：2577-2586.
[10] Dang Z, Watanabe J, Kajino K, et al. Molecular cloning and charac-terization of a T24-like protein in Echinococcus multilocularis[J]. Mol Biochem Parasitol, 2009, 168（1）：117-119.
[11] Ferrer E, Mart nez-Escribano JA, Barderas ME, et al. Peptide epitopes of the Taenia solium antigen Ts8B2 are immunodominant in human and porcine cysticercosis[J]. Mol Biochem Parasitol, 2009, 168（2）：168-171.
[12] Sako Y, Fukuda K, Kobayashi Y, et al. Development of an immuno-chromatographic test to detect antibodies against recombinant Em18 for diagnosis of alveolar Echinococcosis[J]. J Clin Microbiol, 2009, 47（1）：252-254.
[13] Kim SH, Bae YA, Yang Y, et al. Paralogous proteins comprising the 150 kDa hydrophobic-ligand-binding-protein complex of the Taenia solium metacestode have evolved non-overlapped binding affinities toward fatty acid analogs[J]. International Journal for Parasitology, 2011, 41（11）：1207-1215.
[14] Lee EG, Kim SH, Bae YA, et al. A hydrophobic ligand-binding protein of the Taenia solium metacestode mediates uptake of the host lipid：Implication for the maintenance of parasitic cellular homeostasis[J]. Proteomics, 2007, 7（21）：4016-4030.
[15] 许伟.家兔囊虫病的诊断与防治[J].畜牧兽医杂志, 2009, 28（2）：134.
[16] Hopp TP, Woods KR. Prediction of protein antigenic determinants from amino acid sequences[J]. Proc Natl Acad Sci USA, 1981, 78（6）：3824-3828.