AIM:To establish macrophage mannose receiver Model(MMR),and use it to screen active component with mannose receiver(MR)as target from traditional Chinese herbs.METHODS:The mouse abdominal macrophages was hatched with D-mannose and D-galactose of the different concentration,and the flow cytometry and fluorescence microscope were used to measure the antagonistic effect of M-FITC-BSA(Mannose-fluorescein isothiocyanate-bovine serum albumin)with D-mannose and D-galactose.After the MMR screening model was established to screen MR union ingredients of polysaccharide ingredients from six compound prescriptions,such as Siwu Decoction and so on.RESULTS:Both of measuring methods showed that when D-mannose concentration increased the relevance ratio of M? marked with M-FITC-BSA decreases(P

Traditional T vector cloning method requires onerous procedures for identifying recombinant, and directional cloning was impossible. In order to overcome these problems, we have devised a directional T vector pETG based on pET-23a(+). For gene cloning, 7 bp partial LacO sequence was introduced into DNA fragment to reconstitute a full length LacO with Bfu I digested T vector. After transformation, blue colonies were selected on LB plate supplemented with X-gal. Restriction enzyme digestion and PCR identification showed that all blue colonies contained the directionally inserted recombinants and the recombinant efficiency was nearly 100%. We have successfully cloned 103 genes from human liver cDNA; in the study complicated procedures for screening of recombinant were not required. Eight pETG clones were picked for protein expression, and all the clones successfully produced corresponding proteins. We demonstrated that the directional T vector was successfully constructed, and it was very suitable for gene cloning and expression.
Cloning, Molecular ; methods ; DNA, Complementary ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; genetics ; Genetic Vectors ; genetics ; Humans ; Liver ; chemistry ; Polymerase Chain Reaction ; methods ; Recombinant Proteins ; biosynthesis ; genetics