Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (18): 3793-3799.doi: 10.3864/j.issn.0578-1752.2013.18.007

• PLANT PROTECTION • Previous Articles     Next Articles

Molecular Cloning and Expression of Translationally Controlled Tumor Protein of Spodoptera litura (Fab.)

 LU  Meng-Ling, YAN  Chao, MAO  Gen-Lin, XU  Han-Hong   

  1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources/Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou 510642
  • Received:2013-03-18 Online:2013-09-15 Published:2013-05-09

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Abstract: 【Objective】 The objective of this study is to clone the full-length cDNA of TCTP from Spodoptera litura cells, as well as to analyze its sequence.【Method】The full-length cDNA of TCTP was obtained by RACE-PCR. A phylogenetic tree of TCTP protein in different species was constructed. TCTP was cloned in pET-32a (+) and was expressed as fusion protein in Escherichia coli BL21(DE3) cells.【Result】The full-length cDNA of SLTCTP (GenBank accession number: HQ896486.1) was 829 bp with an open reading frame of 519 and encodes 172 amino acids. According to Protparam on-line tools, the deduced protein was predicted to have a computed molecular mass of 19.67 kD. TCTP cloned in pET-32a (+) was expressed as fusion protein in E. coli BL21 (DE3) cells. In order to obtain much target protein of TCTP, optimal culture conditions of pET-32a (+)-TCTP were studied. Induced by 1mmol•L-1 IPTG for 4 h was the best optimized condition for recombinant protein expression. TCTP gene belonged to the TCTP protein family and had a high similarity with other species.【Conclusion】This study has provided a basis of method for further research on the function of SLTCTP and pest control.

Key words: Spodoptera litura , translationally controlled transcription proteins (TCTP) gene , gene clone and expression , sequence analysis

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