Gene screening of dermal papilla cells in the state of aggregative growth and full-length cloning of HSPC016 gene for bioinformatics

Abstract

To screen the genes of dermal papilla cells (DPC) related to the property of aggregative growth, and clone the full-length cDNA of differential HSPC016 gene for functional analysis. DPC were collected from the hair of an individual aged 18 approximately 30 6 hours after the death. The complete papillae of hair were isolated and then cultured. The total DNA was extracted. Suppression subtractive hybridization-polymerase chain reaction was employed to screen out the genes differentially expressed in the DPC under the state of aggregative growth pattern in vitro. Then, rapid amplification of cDNA ends (RACE) technique was used to amplify the full-length cDNA of HSPC016 in the DPC, and bioinformatic methods were used to analyze their possible function. A subtractive library of human DPC was set up, and some up-regulated and down-regulated genes in the DPCs were screened out successfully. HSPC016 was identified to be a gene of 400 bp cDNA. Bioinformatic analyses and databases searching on the Internet indicated that this gene was mapped on chromosome 3 q21.31 and included an open reading frame with 195 bp coding for an expected 64aa soluble protein. The putative protein belonged to PD053992 family and was homologous to T2FA gene in domain. The establishment of subtractive library of DPC provides a solid foundation for further screening the genes related to aggregative growth and analyzing their regulatory mechanisms in DPC. Further more, HSPC016 in DPC may act as a subunit of a functional complex and play a role on transcriptional regulation within nucleus.