Cloning and Expression Analysis of the Mouse Fallotein cDNA Encoded a Vascular Endothelial Growth Factor-like Sequence
| Autor: | Yow-Heng Chen, 陳侑亨 |
|---|---|
| Rok vydání: | 1999 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 87 Delicate tuning of the cell proliferation and differentiation processes of multi-cellular organism depends on the accurate and balanced gene expression regulatory mechanism. Certain genes must be expressed at specific spatial and temporal locations to accomplish the normal development of an organism. Growth factors like epidermal growth factor、transforming growth factor、vascular endothelial growth factor and growth differentiation factors have been known to have profound influence on the regulation of the growth and development of the animal. A novel gene named fallotein has been identified recently in our lab. from human oviduct by PCR technique using the primer pairs designed according to the nucleotide sequences of epidermal growth factor. The cDNA of the novel gene was cloned and named fallotein. The fallotein gene has been proved to be expressed in ovary and uterus. Mouse fallotein PCR primers pairs was designed based on the mouse EST entries homologus to the human fallotein cDNA nucleotide sequences. Through PCR analysis, a 700 bp fallotein cDNA fragment was found, then unitized as probe to clone the full-length cDNA of mouse fallotein from an ovarian cDNA library. The cDNA was 2,692 bp in length as determined by restriction map and nucleotide sequencing, from the nucleotide numbered 198 through 1,235 was a putative protein coding region, that encodes a protein of 345 amino acids in length, 38,741 Da in molecular weight. The first fifteen amino acid residues of the protein complied with the characteristics of the signal peptides of the secretory proteins, from amino acid residues 55 to 160 resemble the CUB domain described by Bork and Beckmann, The amino acid residues at the C-terminal from 250 to 337 was found most similar to the VEGF. The expression pattern of fallotein gene in mouse tissue including : brain、intestine、kidney、liver、lung、ovary、testis and uterus was studied. The resulted will set up the foundation for the exploration of the physiological function of mouse fallotein. The Northern blot analysis performed on the RNA extracted from brain、kidney、liver、lung、ovary、testis and uterus by using the protein coding sequence of fallotein as probe has proved that, the fallotein transcript can be found in all the tissues studied except kidney. The expression level of fallotein was most significant in the lung and the reproductive systems including: ovary、 testis and uterus. The protein expression level of the tissues was found to be in the similar level as the Northern blotting results. In conclusion, the research showed that the N-terminus and the C-terminus of the fallotein protein contains a CUB and VEGF domain seperately, The expression level of the gene was most abundant in the reproduction system like ovary、uterus and testis. Base on these facts, we assumed that the fallotein could be highly correlated to the normal function of the reproductive tracts and gonads, probably through an unknown growth-regulatory mechanism. We believe the correlation deserved further investigation. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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