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Preliminary Study on the Mechanisms for Nuclear Translocation of KGF-2 and bFGF

Author: BaiBingXue
Tutor: FanMing;XuDongGang
School: PLA Military Academy of Medical Sciences
Course: Cell Biology
Keywords: Keratinocyte growth factor-2 Basic fibroblast growth factor Interacting protein Yeast two-hybrid Mammalian cell two-hybrid
CLC: Q78
Type: Master's thesis
Year: 2004
Downloads: 97
Quote: 0
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KGF-2, bFGF nuclear translocation of the preliminary study of the mechanism of KGF-2 and bFGF belong to the fibroblast growth factor family , they are performance cytokines play a role in many important physiological processes . These processes include cell differentiation, proliferation , angiogenesis , tumorigenesis , and damage repair, embryonic development . About they play a role in the molecular mechanisms are not very clear , according to the principles of molecular biology , we combine recombinant yeast two-hybrid two-hybrid , mammalian cells , cell culture method KGF-2 and bFGF proteins interacting with the preliminary research . The findings can be summarized as the following aspects : First, using the PCR method , from a human fetal liver cDNA library successfully fish KGF-2 and bFGF gene . Second, KGF-2 and bFGF interacting proteins using the yeast two-hybrid system screening . Construction of KGF-2 and bFGF the bait protein vector pAS2-1-KGF-2 and pAS2-1-bFGF and were transformed into the yeast host Y190 , KGF-2 and bFGF own transcription activating activity was analyzed , the results showed that the KGF- 2 and bFGF does not have its own transcriptional activation . By the fetal liver cDNA library screening to obtain the two interacting proteins and the presence of KGF-2 . Sequence analysis and homology search revealed : they were to unknown protein PR02605 and people ribosomal protein L22 ( RPL22 ) . The literature search revealed the possibility that they are newly discovered and KGF-2 protein interact . RPL22 understanding more , which comprises three domains , namely the N-domain , the I - domain and C- domain , a classical nuclear localization signal in the N-terminal of RPL22 . The interaction between KGF-2 and RPL22 its way into the nucleus . Third, the use of mammalian cell two-hybrid system in COS-7 cells verified the interaction between the KGF-2 and the human ribosomal protein L22 . Fourth, the use of exogenous KGF-2 and bFGF stimulate NIH/3T3 cells , changes in gene expression observed human ribosomal protein L22 . The results show that KGF- 2 and bFGF acting on cells RPL22 gene expression levels .

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