Cloning, expression, and characterization of three ribosomal protein S13 genes in Sophora flavescens

Authors

  • Yi Liao Anhui University of Traditional Chinese Medicine
  • Miao Miao Liu Anhui University of Traditional Chinese Medicine
  • Qing Ling Li Anhui University of Traditional Chinese Medicine
  • Jun Chu Anhui University of Traditional Chinese Medicine
  • Jing Jing Su Anhui University of Traditional Chinese Medicine
  • Jia Wen Wu Anhui University of Traditional Chinese Medicine

DOI:

https://doi.org/10.14393/BJ-v34n5a2018-39843

Keywords:

Bioinformatics analysis, Gene clone, Ribosomal Protein S13, Sophora flavescens

Abstract

To characterize the structure and function of ribosomal protein S13 (RPS13), we identified full-length open reading frames (ORFs) of three RPS13 genes (RPS13-1, RPS13-2, and RPS13-3) of the Chinese medicinal plant, Sophora flavescens. The target genes were amplified by reverse transcription-polymerase chain reaction (RT-PCR), ligated into the pET22b(+) vector, and then transformed into Escherichia coli BL21 competent cells for protein expression. The physicochemical properties, protein motif, evolution, and structural organization of the three RPS13 genes were analyzed using bioinformatics tools. The full-length ORFs (453 bp) of the three RPS13 genes of S. flavescens were cloned, and each encodes a protein of 151 amino acids in length, and their expression was detected by Western blotting. Bioinformatics analysis showed that RPS13s are stable proteins that are closely related to the 40S RPS13s of Vigna radiate var. radiate. Their three-dimensional structures included three α-helices at the C-terminal and four α-helices at the N-terminal, and the two clusters of helices were connected by a long random coil, which may help maintain the dynamic bridging interactions between the large and small subunits of the ribosome. The full-length ORFs of three RPS13 genes of S. flavescens were successfully cloned and expressed in vitro. The study of the physicochemical properties, evolution, and secondary and three-dimensional structures of the three proteins will provide the theoretical basis for further studies on the function of RPS13s in plants.

Downloads

Download data is not yet available.

Downloads

Published

2018-10-11

How to Cite

LIAO, Y., LIU, M.M., LI, Q.L., CHU, J., SU, J.J. and WU, J.W., 2018. Cloning, expression, and characterization of three ribosomal protein S13 genes in Sophora flavescens . Bioscience Journal [online], vol. 34, no. 5, pp. 1379–1391. [Accessed26 May 2022]. DOI 10.14393/BJ-v34n5a2018-39843. Available from: https://seer.ufu.br/index.php/biosciencejournal/article/view/39843.

Issue

Section

Biological Sciences