Our studies show that DGS provides a kilobase resolution for studying genome structure with

high specificity and high genome coverage. DGS can be applied to validate genome assembly, to compare genome similarity and variation in normal populations, and to identify genomic abnormality including insertion, inversion, deletion, translocation, and amplification in pathological genomes such as cancer genomes.”
“Commercially available bromelain is prepared by performing a tedious and costly purification method that yields bromelain at different degrees of purity. In the current study, a gene encoding NSC 136476 stem bromelain from Ananas comosus was amplified using polymerase chain reaction. This bromelain gene was initially cloned into the pENTR/TEV/D-TOPO vector before being sub-cloned into the pDEST17 expression vector. DNA sequencing of the amplified products selleck chemical exhibited a high level of homology to the corresponding gene from the NCBI public database. Protein expression was conducted in the BL21-Al Escherichia coli strain. The recombinant bromelain was then purified in a single step using immobilized metal affinity chromatography, specifically a Ni-NTA spin column. The purified recombinant bromelain was detected by Western blotting. In addition,

the purified enzyme exhibited hydrolytic activity towards gelatin and a synthetic substrate, LNPE. The purified recombinant bromelain exhibited optimum activity at pH 4.6 and 45 degrees C. (C) 2011 Published by Elsevier Ltd.”
“BACKGROUND: Ethylene glycol monomethyl ether (EGME) exposure is associated with impaired reproductive function. The primary metabolite of EGME is methoxyacetic acid (MAA), a short-chain fatty acid that inhibits historic deacerylase activity and alters gene expression.\n\nOBJECTIVE: Because estrogen signaling is necessary for normal reproductive function and modulates gene expression, the estrogen-signaling pathway is a likely target for MAA; however, GM6001 manufacturer little

is known about the effects of MAA in this regard.\n\nMETHODS: We evaluated the mechanistic effects of MAA on estrogen receptor (ER) expression and estrogen signaling using in vitro and in vivo model systems.\n\nRESULTS: MAA potentiates 17 beta-estradiol (E(2)) stimulation of an estrogen-responsive reporter plasmid in HeLa cells transiently transfected with either a human ER alpha or ER beta expression vector containing a cytomegalovirus (CMV) promoter. This result is attributed to increased exogenous ER expression due to MAA-mediated activation of the CMV promoter. In contrast to its effects on exogenous ER, MAA decreases endogenous ER alpha expression and attenuates E(2)-stimulated endogenous gene expression in both MCF-7 cells and the mouse uterus.\n\nCONCLUSIONS: These results illustrate the importance of careful experimental design and analysis when assessing the potential endocrine-disrupting properties of a compound to ensure biological responses are in concordance with in vitro analyses.