Kyungpook National University, School of Medicine Department of Immunology
Kim Moonkyu, Kim Young-Hee, Im Sang-Uk
Hwang Sun-Young, Chung Eun-Jung, Kim Do-Won, Kim Jung-Chul
Study Information and Results:
Partial sequencing of randomly selected clones from cDNA libraries of specific tissues or cell types to generate expressed sequence tags (ESTs) has proven to be a rapid and efficient means of discovering genes on a large scale and of providing both quantitative and qualitative information regarding gene expression in a variety of tissues. To gain an insight into the genetic profile of human dermal papilla cells, we have constructed cDNA library from cultured dermal papilla cells and generated an expressed sequence tag (EST) database from them.
We constructed cDNA libraries from cultured human scalp hair dermal papilla cells. The 5′ ends of 1,666 randomly selected plasmid clones, each contains one gene which expressed in dermal papilla cell, were sequenced and searched for sequence homologies in the GenBank DNA data base by BLAST. It generated human dermal papilla cell EST database with an average length of 180 nucleotides. Sequences of 351 clones (21.1 percent) showed no significant similarity to entries in the public databases; 1,229 (73.8%) were identical to known human sequences.
The most abundant mRNAs were for type 1(I) collagen chains and fibronectin (53 and 49 times, respectively). mRNAs for osteonectin, α 2(I) collagen, actin-beta, SM22-alpha, and glyceraldehyde-3- phosphate dehydrogenase. Clones for gamma-actin, a 1(VI) collagen, plasminogen activator inhibitor 1, IGH3, a 2(VI) collagen, and ferritin heavy chain showed some redundancy. Clones corresponding to a total of 1,229 specific known genes were classified according to their function.
The most abundant class of genes expressed in dermal papilla, were those for cell structure and motility, which comprises 366 cDNA clones (29.8%), including type I collagen, type IV collagen, fibronectin, osteonectin, beta-actin, alpha-SM22, gamma-actin, and IGH3.
All of the genes in human dermal papilla EST database are available through our web site at http://hair.knu.ac.kr. We also have constructed hair-specific cDNA microarrays containing 3,066 cDNAs using dermal papilla EST clones. Those dermal papilla cDNA collection and microarray can provide valuable information in the field of hair research.