碩士 / 國立臺灣大學 / 生化科學研究所 / 82 / Mitogen-activated protein kinase kinase (MKK) 是一種具有同時對
Tyrosine 和 Serine/Threonine磷酸化的蛋白激脢， 調控 Mitogen-
activated protein(MAP) kinases 的活性。利用RT-PCR ，在鯉魚卵與肝
臟發現兩種 MKKs 的DNA片段，並進行 cDNA基因庫篩選工作，結果只從鯉
魚肝臟 cDNA 基因庫，選殖得到一個含1971bp長度的鯉魚MKK1 cDNA，可
轉譯一個含397個氨基酸的蛋白質。並將其轉譯區域的cDNA，在大腸菌中
進行基因表現。比較該蛋白質的氨基酸序列，發現鯉魚MKK1與rat MKK1，
rat MKK2，Xenopus MKK， Drosophila MKK ，分別有78%，80%，76%
和58%的氨基酸相同。另外再以DIG隨機引子法製備探針，進行鯉魚染色體
基因庫基因選殖， 結果找到鯉魚 MKK1 和 MKK2兩種基因。經核酸定序和
基因結構分析，發現鯉魚MKK1基因有11個exons和10個introns， 全長7.1
kb。 最大的intron ，長1400bp，最小的intron，只有83bp長，但目前仍
欠缺啟動子區域序列。鯉魚MKK2基因，目前只定完部份核酸序列， 分析
其基因結構，發現該序列包含三個exons和四個introns；其中有一個
intron的分佈位置，與鯉魚 MKK1第二個 intron的位置相同，其餘三個
introns的分佈位置，皆不相同。因此，從兩者基因結構比較，鯉魚MKK1
和MKK2在演化上可能屬於不同來源的基因。另外，比較氨基酸序列，發現
目前已知的鯉魚MKK2與鯉魚 MKK1，只有38.2%的氨基酸相同。因此，在魚
類或哺乳類動物 ， 可能存在類似酵母菌兩種以上的 MAP kinase
cascades，以傳遞外界不同的訊息。
We have cloned a full-length cDNA encoding the mitogen-
activated protein kinase kinase (MKK) from a carp liver cDNA
library.The cDNA contains 1971 bp with a single open reading
frame encoding a 397 amino acid protein which is designated
as carp MKK1 . By comparing with known MKK sequences from
other species, carp MKK1 is 78%,80%,76% and 58% identical to
rat MKK1,rat MKK2,Xenopus MKK and Drosophila MKK. The
structural organization of carp mkk1 gene has also been
determined. Carp mkk1 gene comprises eleven exons and ten
introns and spans 7.1 Kb . Among those ten introns,the
longest is 1.4 Kb whereas the shortest is only 83 bp in
length. There is another mkk gene present in carp ovary
tissues detected by RT-PCR.However, the work to isolate this
kind of cDNA clone from a carp ovary cDNA library is not
successful . Only partial sequences of carp mkk2 conding
region was obtains by alignment and combination of those
sequences from genomic clones and RT-PCR products and
these sequences encode 183 amino acid residues . From amino
acid sequence comparison,carp MKK2 is 95.6% and 38.2% identical
to Xenopus MEK2 and carp MKK1 , respectively . Therefore,it is
possible that there are at least two different MAP kinase
cascades involved in the vertebrate signal transduction pathway.
| Identifer | oai:union.ndltd.org:TW/082NTU00103006 |
| Date | January 1994 |
| Creators | Ming-Shyue Lee, 李明學 |
| Contributors | Tung-Bin Lo, 羅銅壁 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 51 |
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