cDNA cloning and nucleotide sequencing
of human connective tissue growth factor
obtained from primary pulpal fibroblasts
Authors :
- Pasutha Thunyakitpisal D.D.S., Ph.D.
Abstract :
Objective To clone human connective tissue growth factor (hCTGF) from pulpal fibroblasts.
Materials and methods Primary human pulpal fibroblasts were isolated from pulpal tissues.
The reverse transcriptase-PCR assay was used to amplify the cDNA of hCTGF. The amplified PCR
products were ligated into the TOPO R cloning vector and transformed into competent bacteria cells.
The putative clones were bidirectionally sequenced to analyze nucleotide sequence and compare with
hCTGF cDNA sequence references.
Results From RT-PCR reaction, expression of CTGF mRNA was detected in human pulpal
fibroblast. Through bi-directional sequencing analysis, nucleotide sequence of our hCTGF has 100%
homology to the hCTGF sequence reported.
Conclusion Human pulpal fibroblasts express CTGF mRNA. The hCTGF cDNA obtained from
primary pulpal fibroblast has 100% homology to hCTGF sequence references.
(CU Dent J. 2007;30:227-34)
Keywords :
cloning; connective tissue growth factor; pulpal fibroblasts
|
