EH质量流量计培训资料.ppt

华中科技大学硕士学位论文
Abstract
mAmetrine is a yellow fluorescent protein (YFP) with large Stokes shift (LSS). LSS
is an accepted nomenclature in the field of fluorescent proteins to denote fluorescent
proteins exhibiting excitation and emission maxima that differ greater than ~100 nm.
mAmetrine retains the property of violet excitation (excitation wavelength, ex = 406 nm),
and yellow emission (emission wavelength, em = 526 nm) resulting from excited-state
proton transfer (ESPT) and π-π stacking interactions with the chromophore. However, the
mechanism of ESPT for mAmetrine LSS fluorescence emission is unknown. To better
understand mAmetrine function and study ESPT pathway or other pathway(s) responsible
for LSS, the three-dimensional (3-D) structure of mAmetrine must be determined by
performing X-ray crystallography. X-ray crystallography involved a series of steps:
cloning; expression; large-scale purification; crystallization; collection, integration, and
refinement of diffraction data; and determination of atomic positions. The protein
crystallization technique used was hanging drop vapor-diffusion technique. The 3-D
structure of mAmetrine was determined at resolution Å and the data was analyzed to
study the mechanism of ESPT pathway or other pathway(s) responsible for LSS. Further
site-directed mutagenesis (F203Y, S205N and E222K) were required to finalize the
structural analysis of mAmetrine.

Key words:mAmetrine fluorescent protein(s) Large-Stokes Shift (LSS)
protein crystallography X-ray diffraction
excited-state proton transfer (ESPT)
II
华中科技大学硕士学位论文
TABLE OF CONTENTS
摘要............................................................................................................... I
ABSTRACT .................................................................................................... II
CHAPTER 1
Introduction ...................................................................................