Molecular Cloning, Characterization and Expression Analysis of Calreticulin Gene in the Ant Polyrhachis vicina Roger (Hymenoptera: Formicidae)

Liping Liu, Liang Dang, Gengsi Xi, Fang Wang

Abstract


Calreticulin (CRT) as a ubiquitous and highly conserved calcium-binding protein exists in endoplasmic reticulum (ER), which possesses a variety of biological functions in the regulation of cell calcium homeostasis, molecular chaperoning and innate immunity. In our research, the calreticulin homologous gene (refered as PvCRT) was cloned from the ant Polyrhachis vicina Roger (Hymenoptera: Formicidae), the full-length cDNA of PvCRT is 1584bp base pairs(bp), contains a 5’-untranslated region of 87bp and a 3’-untranslated region of 246bp. The open reading frame (ORF) of 1251bp encodes 416 amino acids. Using real-time quantitative RT-PCR to study PvCRT mRNA expression levels indicate that this gene was expressed in different developmental stages of castes of P. vicina. The mRNA expression level in both embryos and adults show that PvCRT gene may play some essential roles in the ant’s development.

Keywords


Calreticulin homologous gene; Molecular cloning; real-time quantitative RT-PCR; Polyrhachis vicina

Full Text:

PDF

References


Bendtsen, J.D., H. Nielsen, G., Von Heijine & S. Brunak (2004). Improved prediction of signal peptides: SignalP 3.0. J. Biochem. Mol. Biol., 40:783-795. doi: 10.1016/j.jmb.2004.05.028

Fitzgerald M. & T. Shenk (1981). The sequence 5’-AAUAAA-3’forms parts of the recognition site for polyadenylation of late SV40 mRNAs. Cell, 24(1): 251-260.

Gamo, S., Dodo, K., Matakatsu, H. & Tanaka, Y. (1998). Molecular genetical analysis of Drosophila ether sensitive mutants. Toxicol. Lett., 23: 100-101:329-337. doi: 10.1016/S0378-4274(98)00203-3

Gamo, S., Tomida, J., Dodo, K., Keyakidani, D., Matakatsu, H. & Yamamoto, D. (2003). Calreticulin mediates anesthetic sensitivity in Drosophila melanogaster. Anesthesiology, 99: 867–875.

Gelebart P., Opas M. & Michalak M. (2005). Calreticulin, a Ca2+-binding chaperone of the endoplasmic reticulum. Int. J. Biochem. Cell Biol., 37: 260-266. doi.org/10.1016/j.biocel.2004.02.030

Guo, X.J. & G.S., Xi (2010). Molecular Cloning and Expression Analysis of the Gene Encoding MEF2 in the Ant Polyrhachis vicina (Hymenoptera:Formicidae). Sociobiology 56: 235-248.

Khanna, N.C., M. Tokuda & Waisman, D.M. (1987) Comparison of calregulins from vertebrate livers. Biochem. J., 242(1):245-251.

Kuraishi, T., Manaka, J., Kono, M., Ishii, H., N. Yamamoto & Koizumi, K. (2007). Identification of calreticulin as a marker for phagocytosis of apoptotic cells in Drosophila. Exp. Cell Res., 313: 500–510. doi: 10.1016/j.yexcr.2006.10.027

Lee M.S. & G.M. Lee .(2001). Effect of hypoosmotic pressure on cell growth and antibody production in recombinant Chinese hamster ovary cell culture. Cytotechnology. 36(1-3):61-9

Lv, S.M., Xi, G.S. & Wang, X..H. (2008). Molecular cloning characterization and expression analysis of a QM homologue in the ant Polyrhachis vicina (Hymenoptera:Formicidae). Can. Entomol., 140: 312-323. doi: 10.4039/n08-009

Luana, W., Li, F., Wang, B., Zhang, X., Liu, Y. & Xiang, J. (2007). Molecular characteristics and expression analysis of calreticulin in Chinese shrimp Fenneropenaeus chinensis. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 147: 482-491. doi: 10.1016/j.cbpb.2007.03.001

Lynch, J.M., Chilibeck, K., Qui, Y. & Michalak, M. (2006). Assembling pieces of the cardiac puzzle: calreticulin and calcium-dependent pathways in cardiac development, health, and disease. Trends Cardiovasc. Med. 16(3):65-69.

Mesaeli, N., Nakamura, K., Zvaritch, E., Dickie, P., Dziak, E., Krause, K.H., Opas, M., MacLennan, D.H. & Michalak, M. (1999). Calreticulin Is Essential for Cardiac Development. J. Cell Biol., 144: 857–868. 10.1083/jcb.144.5.857

Ostwald, T.J. & MacLennan, D..H. (1974). Isolation of a high affinitycalcium-binding protein from sarcoplasmic reticulum. J. Biol. Chem. 249: 974–979.

Park, B.J., Lee, D.G. , Yu, J.R., Jung, S.K., Choi, K., Lee, J., Lee, J., Kim, Y.S., Lee, J.I., Kwon, J.Y., Lee, J., Singson, A., Song, W.K., Eom, S.H., Park, C.S., Kim, D.H.,

Bandyopadhyay, J. & Ahnn, J. (2001). Calreticulin, a Calcium-binding Molecular Chaperone, is Required for Stress Response and Fertility in Caenorhabditis elegans. Mol. Biol. Cell., 12: 2835–2845. doi: 10.1091/mbc.12.9.2835

Rauch, F., Prud’homme, J., Arabian, A., Dedhar, S & St-Arnaud, R. (2000). Heart brain, and body wall defects in mice lacking calreticulin. Exp. Cell Res. 256(1):105-111. doi: 10.1006/excr.2000.4818

Rodriguez-Mdel, C., Martinez-Barnetche, J., Alvarado-Delgado, A., Batista, C., Argotte-Ramos, R.S. & Hernandez-Martinez, S. (2007). The surface protein Pvs25 of Plasmodium vivax ookinetes interacts with calreticulin on the midgut apical surface of the malaria vector Anopheles albimanus. Mol. Biochem. Parasitol., 153: 167–177. doi: 10.1016/j.molbiopara.2007.03.002

Silerová, M., Kauschke, E., Procházková, P., Josková, R., Tucková, L. & Bilej, M. (2007). Characterization, molecular cloning and localization of calreticulin in Eisenia fetida earthworms. Gene, 397(1-2):169-77. doi: 10.1016/j.gene.2007.04.035

Simonet, G., Poels, J., Claeys I., Van Loy, T., Franssens, V., De Loof A. & Vanden Broeck J.) (2004). Neuroendochrinological and molecular aspects of insect reproduction. J. Neuroendocrinol.,16: 649 -659. doi: 10.1111/j.1365-2826.2004.01222.x

Smith M.J. & Koch, G.L. (1989). Multiple zones in the sequence of calreticulin (crp55, calregulin, HACBP), a major calcium binding ER/SR protein. EMBO (Eur. Mol. Biol. Organ.) J., 8: 3581-3586.

Stoltzfus, J.R., Horton, W.J. & Grotewiel, M.S. (2003). Odor-guided behavior in Drosophila requires calreticulin. J. Comp. Physiol., 189: 471–483. doi: 10.1007/s00359-003-0425-z

Tamura, K., Dudley, J., Nei, M. & Kumar, S. (2007). MEGA4: molecular evolutionary geneticsanalysis (MEGA) software version 4.0. Mol. Biol. Evol., 24: 1596–1599. Doi: 10.1093/molbev/msm092

Wang, L, Fang, Q., Zhu, J., Wang, F., Rean Akhtar, Z. & Ye, G. (2012) Molecular cloning and functional study of calreticulin from a lepidopteran pest Pieris rapae. Dev. Comp. Immunol., 38(1): 55-65. doi: 10.1016/j.dci.2012.03.019

Zhang, C.Y., Xu, S.G. & Huang, X.X. (2005). A novel and convenient relative quantitative method of fluorescence real time RT-PCR assay based on slope of standard curve. Prog. Biochem. Biophysiol., 32: 883-888.




DOI: http://dx.doi.org/10.13102/sociobiology.v60i4.355-361

Refbacks

  • There are currently no refbacks.


JCR Impact Factor 2018: 0.604