| Project/Area Number |
08457467
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Ophthalmology
|
|---|
| Research Institution | Kyoto Prefectural University of Medicine |
|---|
Principal Investigator |
KINISHITA Shigeru Kyoto Prefectural University of Medicine Medicine, Ophthalmology, Professor, 医学部, 教授 (30116024)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
OKUBO Kousaku Kyoto Prefectural University of Medicine Molecular, and Cellular Biology, Osaka, 細胞生体工学センター, 助教授 (40233069)
NISHIDA Kohji Kyoto Prefectural University of Medicine Medicine, Ophthalmology, Instructor, 医学部, 助手 (40244610)
|
|---|
| Project Period (FY) |
1996 – 1997
|
| Project Status |
Completed (Fiscal Year 1997)
|
| Budget Amount *help |
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1997: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1996: ¥4,600,000 (Direct Cost: ¥4,600,000)
|
|---|
| Keywords | corneal epithelium / cDNA analysis / human / cDNA library / specific genes / keratin 12 / cathepsin |
|---|
| Research Abstract |
We analyzed the composition of mRNA in human corneal epithelium. From these data, we selected unknown cornea-specific genes, cloned the full-lengh cDNA,found the chromosomal localization and analyzed the functions of genes.
A full-lenght cDNA library was constructed from poly A+RNA of human corneal epithelium collected at ophthalmic surgeries. Using the library, positive clones were screened with fragments of 3'-directed EST.Inserted cDNA of these clones was sequenced and analyzed. As a result, we isolated three novel genes. First, cornea-specific human keratin 12 encoding 498 amino acids was isolated. It was found that it mapped to 17q12 and was composed of 8 exons and 7 introns. Keratin 12 mutations were detected in Meesmann's corneal dystrophy. Second, a novel cathepsin cDNA which contained ORF 77% identical homology to human cathepsin L was found. A recombinant protein produced using a baculovirus expression system has proteolytic activity that characterizes it as a cysteine proteinase. These facts show tthat the genes are near relatives, therefore, we named the novel gene cathepsin V.The expression of cathesin V was examined in tissues by RT-PCR.Only in cornea, the expression level was higher than that of human cathepsin L.Cathepsin V may play an important role in corneal physiology. A third gene, unique to corneal epithelium, was identified as a member of the family of four transmembrane proteins by homology analysis.
|
