碩士 / 國立臺灣大學 / 生化科學研究所 / 82 / CDC2蛋白激脢的活化與高等動物細胞週期的分裂期進行有關。而在動物組
織中，已知至少存在六種結構和 CDC2相似的CDK分子，配合著不同的
cyclin擔任調節細胞週期的不同角色。利用RT-PCR在鯉魚組織找到數種類
似cdk的DNA片段，並在卵巢組織 cDNA 基因庫篩選出一個全長1.3Kb的
cDNA clone，可轉譯一個含 302個氨基酸的CDC2 蛋白激脢，此蛋白激脢
和各種真核生物的CDC2比較，相同的氨基酸約佔80%。為了研究 CDC2的基
因調控，我們進一步從鯉魚染色體基因庫中篩選出含 cdc2基因的phage
clone，並定出長約 5Kb 的部份DNA序列，此序列涵蓋cdc2基因的啟動子
區域和cdc2基因的蛋白質轉譯區，分析 cdc2 基因的啟動子區域DNA 序列
，發現存在多種和基因轉錄有關的核酸序列，如：和細胞週期有關的E2F
，p53結合序列，和左右生長分裂或細胞分化的 c-Myb結合序列，以及與
急性狀態反應有關的 NF-IL6， H-APF-1，HNF-5結合序列。另外利用RT-
PCR在鯉魚不同組織也找到類似cdk6的 DNA片段，經核酸定序發現來自脾
臟的 Sp82 植株其序列與來自卵巢組織的 Ov153植株完全相同，但多出
87bp 。利用 Sp82 所標定之核酸探針，也從鯉魚染色體基因庫中篩選出
含鯉魚 cdk6基因的 phage clone。 完全定出長5kb的部份DNA序列後，發
現Sp82植株所多出的87bp屬於intron的序列，因此這87bp可能由於
alternative splicing 所造成，或由 mRNA imcomplete processing 所
造成。比較鯉魚cdc2，cdk6和人類的cdc2基因結構，發現它們的intron
splice sites很類似，但intron大小不同，如此cdc2和cdk6基因可能是由
同一古老的基因演化而來。
Cyclin-dependent kinases (CDKs) control eukaryotic cell cycle
progression. CDK must bind to respective cyclin to gain
its kinase activity. CDK activities also depends
on its phosphorylation state. P34(cdc2) is the archetype
of the CDK family and controls entry into mitosis in all
eukaryotes. Other distant CDKs, such as cdk2,cdk4 and cdk5,
seem to control G1 and S phase functions. To study the
transcriptional regulation of these cdk genes, we reported
the genomic structure of carp cdc2 and cdk6 genes. Several cdk-
related DNA fragments were amplified by RT-PCR. Using these
DNA as probe, we isolated a 1.3 kb clone from carp ovary cDNA
library encoding 302 amino acid residues. Comparing with
other known CDC2 sequences, carp CDC2 is 82%, 81%, 83.5%, 69%,
an 62.6% identical to human CDC2, chicken CDC2, frog CDC2,
fruit fly CDC2 and fission yeast CDC2, respectively. The
gonomic structure of carp cdc2 was determined. DNA fragment
covering about 2kb promoter region and most cdc2 ccoding
region. We find many kinds of mammalian transcription factor
binding sequences in the promoter region, including (1) E2F,
p53-binding sequences relevant to cell cycle control; (2) c-
Myb binding sequences playing roles in cell proliferation and
differentiation and (3) NF-IL6, H-APF-1, HNF-5 binding
sequences which response to acute phase stimulation. In RT-PCR,
we found that Sp82 from spleen are the same as Ov153 from ovary
except for one insertion of 87bp. We isolated the genomic
clones containing the Sp82. After sequencing about 5kb
genomic DNA, we found that the 87bp insertion in Sp82 are
the intron sequences .It seems that tisue specific
alternative splicing may be present in the prossing of carp
cdk6 mRNA.
| Identifer | oai:union.ndltd.org:TW/082NTU00103004 |
| Date | January 1994 |
| Creators | Hsiao,Pei-Wen, 蕭培文 |
| Contributors | Chang,Geen-Dong;Huang,Chang-Jen, 張震東;黃銓珍 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 50 |
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