Molecular cloning, characterization and localization of chicken type II procollagen gene

• Published in: Gene 2006-01, Vol.366 (1), p.67-76
• Main Authors: Xi, Caixia, Liu, Nan, Liang, Fei, Guo, Siqi, Sun, Yuying, Yang, Fengtang, Xi, Yongzhi
• Format: Article
• Language: English
• Subjects: 3' Flanking Region - genetics; 4′-6′-diamino-2-phenylindole; 5' Flanking Region - genetics; Administration, Oral; Amino Acid Sequence; amino-terminal propeptide; Animals; Arthritis, Rheumatoid - drug therapy; base pair; C-propeptide; carboxyl-terminal propeptide; ccol2a1; chicken alpha 1 type II collagen; Chickens - genetics; Chondrogenesis - physiology; Cloning, Molecular - methods; Collagen Type II - administration & dosage; Collagen Type II - biosynthesis; Collagen Type II - genetics; DAPI; Developmental dynamics; DNA, Complementary - genetics; Evolutional characters; Exons - genetics; Expression; FISH; FITC; fluorescein isothiocyanate; fluorescence in situ hybridization; Gallus gallus domesticus; Gene Expression Regulation - physiology; Genomic organization; Humans; kilobases; Molecular Sequence Data; N-propeptide; Organ Specificity - genetics; Protein Precursors - administration & dosage; Protein Precursors - genetics; Protein Sorting Signals - genetics; Protein Structure, Tertiary - genetics; RACE; rapid amplification of cDNA ends; reverse transcription and polymerase chain reaction; RT-PCR; SOE-PCR; Splicing; splicing by overlapping extension PCR; untranslated region; UTR
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2005.06.032

Chicken type II procollagen ( ccol2a1) has become as an important oral tolerance protein for effective treatment of rheumatoid arthritis. However, its molecular identity remains unclear. Here, we reported the full-length cDNA and nearly complete genomic DNA encoding ccol2a1. We have determined the structural organization, evolutional characters, developmental expression and chromosomal mapping of the gene. The full-length cDNA sequence spans 4837 bp containing all the coding region of the ccol2a1 including 3′ and 5′ untranslation region. The deduced peptide of ccol2a1, composed of 1420 amino acids, can be divided into signal peptide, N-propeptide, N-telopeptide, triple helix, C-telopeptide and C-propeptide. The ccol2a1 genomic DNA sequence was determined to be 12,523 bp long containing 54 exons interrupted by 53 introns. Comparison of the ccol2a1 with its counterparts in human, mouse, canine, horse, rat, frog and newt revealed highly conserved sequence in the triple helix domain. Chromosomal mapping of ccol2a1 locates it on 4P2. While the ccol2a1 mRNA was expressed in multiple tissues, the protein was only detected in chondrogenic cartilage, vitreous body and cornea. The ccol2a1 was found to contain two isoforms detected by RT-PCR. The distribution of the ccol2a1 lacking exon 2wasfrequently detected in chondrogenic tissues, whereas the exon 2-containing isoform was more abundant in non-chondrogenic tissues. These results provide useful information for preparing recombinant chicken type II collagen and for a better understanding of normal cartilage development.