Cloning, Characterization and Bioinformatic Analysis of the Gene Encoding the Larval Serum Protein 2 in Diapause of the Onion Maggot, Delia Antiqua

Jingjing Xu, Bin Chen, Zhengbo He, Youjin Hao

Institute of Entomology and Molecular Biology, College of Life Sciences, Chongqing Normal University, Shapingba, Chongqing, China.
nstitute of Entomology and Molecular Biology, College of Life Sciences, Chongqing Normal University, Shapingba, Chongqing, China.

DOI: 10.4236/eng.2013.510B100                

Abstract

The full-length cDNA encoding Larval serum protein 2 (LSp-2) in the onion maggot,Delia antiqua, was cloned and sequenced by rapid ampli?cation of cDNA ends methods. The result showed that the cDNA was 2203 bp long and the open reading frame (ORF) of 2106 bp encoded 701 amino acid with a calculated molecular weight of 80.5 kDa and an isoelectric point of 5.87. The onion maggot LSp-2 shows highest homology (83%) to that ofCalliphora vicinaat amino acid level. Its signal peptides, domains and structures were predicted and analyzed by using bioinformatic methods. The amino acid sequence of LSP-2 suggests that it would be a typical hexamerin.

Keywords

Delia Antiqua; Diapause; Larval Serum Protein 2; Cloning; Molecular Characterization; Bioinformatic Analysis

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	S. Mousseron-Grall, J. Kejzlarova-Lepesant, T. Burmester and C. Chihara, “Sequence, Structure and Evolution of the Ecdysone-Inducible Lsp-2 Gene of Drosophila melanogaster,” Biochemistry, Vol. 245, 1997, pp. 192- 198.
[2]	S. O. Zakharkin, A. V. Gordadze and S. E. Korochkina, “Molecular Cloning and Expression of a Hexamerin cDNA from the Malaria Mosquito, Anopheles gambiae,” Biochemistry, Vol. 246, 1997, pp. 719-726.
[3]	T. Burmester, C. Antoniewski and J.-A. Lepesant, “Ecdysone-Regulation of Synthesis and Processing of Fat Body Protein 1, the Larval Serum Protein Receptor of Drosophila melanogaster,” Biochemistry, Vol. 262, 1999. pp. 49-55.
[4]	B. Chen and T. Kayukawa, “DaTrypsin, a Novel Clip- Domain Serine Proteinase Gene Up-Regulated during Winter and Summer Diapauses of the Onion Maggot,” Delia antiqua,” Gene, Vol. 347, 2005, pp. 115-123. http://dx.doi.org/10.1016/j.gene.2004.12.026
[5]	D. A. Hahn1, L. N. James1 and K. R. Milne1, “Life History Plasticity after Attaining a Dietary Threshold for Reproduction Is Associated with Protein Storage in Flesh Flies,” NIH Public Access, Vol. 22, No. 6, 2008, pp. 1081-1090.
[6]	C. K. Moreira, M. de L. Capurro and M. Walter, “Primary Characterization and Basal Promoter Activity of Two Hexamerin Genes of Musca domestica,” Journal of Insect Science, Vol. 4, 2004, p. 2.
[7]	A. S. Cristino and F. M. F. Nunes, “Organization, Evolution and Transcriptional Profile of Hexamerin Genes of the Parasitic Wasp Nasonia vitripennis (Hymenoptera: Pteromalidae),” Insect Molecular Biology, Vol. 19, 2010. pp. 137-146. http://dx.doi.org/10.1111/j.1365-2583.2009.00970.x
[8]	J. Godlewski, B. Kludkiewicz and K. Grzelak, “Expression of Larval Hemolymph Proteins (Lhp) Genes and Protein Synthesis in the Fat Body of Greater Wax Moth (Galleria mellonella) Larvae during Diapause,” Journal of Insect Physiology, Vol. 47, 2001, pp. 759-766. http://dx.doi.org/10.1016/S0022-1910(01)00050-6?
[9]	P. Nagamanju, I. A. Hansen and T. Burmester, “Complete Sequence, Expression and Evolution of Two Members of the Hexamerin Protein Family during the Larval Development of the Rice Moth, Corcyra cephalonica,” Insect Biochemistry and Molecular Biology, Vol. 33, 2003, pp. 73-80. http://dx.doi.org/10.1016/S0965-1748(02)00178-9
[10]	C. A. D. De Korta and A. B. Koopmanschapa, “Nucleotide and Deduced Amino Acid Sequence of a cDNA Clone Encoding Diapause Protein 1, an Arylphorin-Type Storage Hexamer of the Colorado Potato Beetle,” Journal of Insect Physiology, Vol. 40, 1994, pp. 527-535. http://dx.doi.org/10.1016/0022-1910(94)90126-0
[11]	J. Tungjitwitayakul, T. Singtripop, A. Nettagul, Y. Oda, N. Tatun, T. Sekimoto and S. Sakurai, “Identification, Characterization, and Developmental Regulation of Two Storage Proteins in the Bamboo Borer Omphisa fuscidentalis,” Journal of Insect Physiology, Vol. 54, 2008, pp. 62-76.