日本人做生物还真牛~~

且不说我的领域的SSH/LIMK都是是日本人发现的。
这篇science加上两年前的EMBO，用精细的显微镜分析加上green-red转化的荧光蛋白，巧妙分析了细胞分裂早期的现状，牛！

Science 4 May 2007:
Vol. 316. no. 5825, pp. 719 – 723
DOI: 10.1126/science.1138591
Blastocyst Axis Is Specified Independently of Early Cell Lineage But Aligns with the ZP Shape
Yoko Kurotaki,1,2* Kohei Hatta,3{dagger} Kazuki Nakao,4 Yo-ichi Nabeshima,1,2 Toshihiko Fujimori1,2{ddagger}

The mechanisms controlling the establishment of the embryonic-abembryonic (E-Ab) axis of the mammalian blastocyst are controversial. We used in vitro time-lapse imaging and in vivo lineage labeling to provide evidence that the E-Ab axis of the mouse blastocyst is generated independently of early cell lineage. Rather, both the boundary between two-cell blastomeres and the E-Ab axis of the blastocyst align relative to the ellipsoidal shape of the zona pellucida (ZP), an extraembryonic structure. Lack of correlation between cell lineage and the E-Ab axis can be explained by the rotation of the embryo within the ZP.

EMBO reports 6, 3, 233–238 (2005)
DOI: 10.1038/sj.embor.7400361
Semi-rational engineering of a coral fluorescent protein into an efficient highlighter
Hidekazu Tsutsui1, Satoshi Karasawa1, 3, Hideaki Shimizu2, Nobuyuki Nukina2 & Atsushi Miyawaki1
1 Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan
2 Laboratory for Structural Neuropathology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan
3 Amalgaam Co., Ltd, 2-9-3 Itabashi, Itabashi-ku, Tokyo 173-0004, Japan

Kaede is a natural photoconvertible fluorescent protein found in the coral Trachyphyllia geoffroyi. It contains a tripeptide, His 62-Tyr 63-Gly 64, which acts as a green chromophore that is photoconvertible to red following (ultra-) violet irradiation. Here, we report the molecular cloning and crystal structure determination of a new fluorescent protein, KikG, from the coral Favia favus, and its in vitro evolution conferring green-to-red photoconvertibility. Substitution of the His 62-Tyr 63-Gly 64 sequence into the native protein provided only negligible photoconversion. On the basis of the crystal structure, semi-rational mutagenesis of the amino acids surrounding the chromophore was performed, leading to the generation of an efficient highlighter, KikGR. Within mammalian cells, KikGR is more efficiently photoconverted and is several-fold brighter in both the green and red states than Kaede. In addition, KikGR was successfully photoconverted using two-photon excitation microscopy at 760 nm, ensuring optical cell labelling with better spatial discrimination in thick and highly scattering tissues.

只是KikGR的体内转变需要使用two-photon，唉，短期内是实验不了了