The discovery of the interaction between augmenter of liver regeneration and Na~+, K~+-ATPase with yeast two-hybrid system / 中国病理生理杂志

AIM: To screen the proteins interacting with human augmenter of liver regeneration (hALR) by yeast two-hybrid system and to study the mechanism of hALR action. METHODS: hALR bait plasmid was constructed by ligating the gene of hALR into pGBKT7, then transformed into yeast AH109. The yeast strain AH109 containing pGBKT7-hALR was mated with yeast Y187 containing human liver cDNA library plasmid. Diploid yeast was plated on SD/-trp-leu-his-ade (QDO) for screening and on QDO containing X-?-gal for further selection.The AD/library inserts were amplified by PCR and the PCR products were characterized by digesting with Sau3AⅠ and HaeⅢ restriction enzyme to eliminate the duplicates. After sequencing, the positive clones were analysed by bioinformatics. RESULTS: Several positive clones were obtaind. The sequencing and analysis shown that one of them is 669 bp DNA fragment encoding ? subunit of Na +,K +-ATPase. The 224 bp 3′terminal DNA fragment is non-encoder region, and the 445 bp 5′terminal DNA encodes C-terminal 147 amino acid residues of Na +, K +-ATPase ? subunit. CONCLUSION: The results of screening proteins using yeast two-hybrid system showed that hALR could interact directly with Na +, K +-ATPase in the yeast cell.

Advances in the research of bFGF binding to heparin / 中国病理生理杂志

Basic fibroblast growth factor (bFGF) is a multi-potential growth factor whose biological activities depend on its receptor's intrinsic tyrosine kinase activity and second messengers such as the mitogen activated protein kinases. Heparin sulfate proteoglycans (HSPGs) have been demonstrated to enhance or inhibit bFGF activity. The response elicited by HSPG is related to the relative concentrations and binding kinetics for bFGF of the various pools of HSPG. The type of cellular response might depend on the specific HSPG and FGF receptor expressed on the cell surface. The specific core protein of HSPG, and tissue specific differences in heparin sulfate modification result in altered bFGF regulation.

STUDY OF PROBE-PREPARATION OF THE EXPRESSION GENES FOR K562 CELL LINE / 解剖学报

Objective To develop a better probe preparation method of gene chips for the further gene chips fabrication and its application in the gene expression. Method Using human leukemia K562 cells as material,applying a new kind of technology for gene differential display restriction display(RD PCR),we isolated a lot of cDNA fragments as probes of gene chips. Results This method has overcome the shortcoming of more pseudopositives in DD PCR,and was easy to be manipulated.Compared with the chip fabricated with full cDNA probes,the probe length is comparatively homogeneous,and the hybridization dynamics is apt to be controlled.Conclusion\ RD PCR technology could provide a new strategy for the probe preparation of gene chips.\;[