Molecular Characterization, Bioinformatic Analysis, and Expression Profile of Lin-28 Gene and Its Protein from Arabian Camel ( Camelus dromedarius )

• Published in: International journal of molecular sciences Vol. 20; no. 9; p. 2291
• Main Authors: Alharbi, Sultan N, Alduhaymi, Ibtehal S, Alqahtani, Lama, Altammaami, Musaad A, Alhoshani, Fahad M, Alrabiah, Deema K, Alyemni, Saleh O, Alsulami, Khulud A, Alghamdi, Waleed M, Fallatah, Mohannad
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 09.05.2019; MDPI
• Subjects: 3D structure; Amino Acid Sequence; Amino acids; Animals; Arabian Camel; Base Sequence; bioinformatics analysis; Camels; Camelus - genetics; Camelus dromedarius; cold-shock domain (CSD); Computational Biology - methods; Deoxyribonucleic acid; DNA; Electrophoresis; Flexibility; Gel electrophoresis; Gene expression; Gene Expression Profiling; Gene Expression Regulation; Genes; Genomes; Lin-28; Mammals; Mass spectrometry; Mass spectroscopy; MicroRNAs; Models, Molecular; Molecular weight; Nucleotide sequence; Peptides; Peptides - chemistry; phylogenetic analysis; Phylogeny; PMF-MS; Protein Structure, Secondary; Proteins; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA-Binding Proteins - chemistry; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; Scientific imaging; Search engines; Spectroscopy; Stem cells; Therapeutic applications; phylogenetic analysis; Arabian Camel; cold-shock domain (CSD); Lin-28; 3D structure; PMF-MS; bioinformatics analysis
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms20092291

Lin-28 is an RNA-binding protein that is known for its role in promoting the pluripotency of stem cells. In the present study, Arabian camel Lin-28 (cLin-28) cDNA was identified and analyzed. Full length cLin-28 mRNA was obtained using the reverse transcription polymerase chain reaction (RT-PCR). It was shown to be 715 bp in length, and the open reading frame (ORF) encoded 205 amino acids. The molecular weight and theoretical isoelectric point (pI) of the cLin-28 protein were predicted to be 22.389 kDa and 8.50, respectively. Results from the bioinformatics analysis revealed that cLin-28 has two main domains: an N-terminal cold-shock domain (CSD) and a C-terminal pair of retroviral-type Cysteine3Histidine (CCHC) zinc fingers. Sequence similarity and phylogenetic analysis showed that the cLin-28 protein is grouped together and . Quantitative real-time PCR (qPCR) analysis showed that cLin-28 mRNA is highly expressed in the lung, heart, liver, and esophageal tissues. Peptide mass fingerprint-mass spectrometry (PMF-MS) analysis of the purified cLin-28 protein confirmed the identity of this protein. Comparing the modeled 3D structure of cLin-28 protein with the available protein 3D structure of the human Lin-28 protein confirmed the presence of CSD and retroviral-type CCHC zinc fingers, and high similarities were noted between the two structures by using super secondary structure prediction.