| Gene Expression Profile of Mouse Calvarial Osteoblast by cDNA Library |
| cDNA Library를 이용한 생쥐 두개골 유래 조골세포의
유전자 발현 분석 |
| 정연관,정재환,최제용 |
| 경북대학교 의과대학 생화학•세포생물학교실 |
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| Abstract |
Background
Large-scale sequencing of cDNA clones from cDNA library is a powerful approach for the discovery of novel genes, identification of novel gene family members and definition of gene expression profiles. Listing up of bone related genes is necessary for better understanding of molecular bone physiology and homeostasis.
Methods
Primary cultured mouse calvarial osteoblast (MCO) was isolated from mouse embryo at day 17.5. Total RNA was purified from MCO and constructed cDNA library using Stratagen cDNA library contstruction kit. Random selected clones of cDNA library were purified and analyzed the sequence by sequencing analysis. Sequences were identified using BlastN in NCBI mouse EST database.
Results
The high quality of MCO cDNA library was constructed showing low background (7.2%) such as no insert clone, mitochondrial DNA, E. coli gene, etc. Total 2361 clones were identified by random sequencing. Except low sequencing quality (207), 2154 sequences were further analyzed. In analyzed 1999 useable clones, 1137 clones were known gene, 231 clones were similar to known genes, 69 clones were novel genes, and 562 clones were expression sequence tag (EST). Majority of identified 1078 clones were detected only once, and the others were over two times. The most abundant genes were type I collagen, α2(I) and α1(I) that were detected 36 and 32 times each. Also osteopontin, actin, Thbs1, Fth and Lgals1 were detected highly.
Conclusion
In NCBI EST library database, there is only one library related to mouse osteoblasts. Our results will provide a framework data for understanding the gene expression in osteoblasts.
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| Key Words:
cDNA library, Expression sequence tag (EST), Mouse calvarial osteoblast, BLASTN, Type I collagen |
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