The human genome holds an extraordinary trove of information about human development, physiology, medicine and evolution. Here we report the results of an international collaboration to produce and make freely available a draft sequence of the human genome. We also present an initial analysis of the data, describing some of the insights that can be gleaned from the sequence.

To illuminate the function and evolutionary history of both genomes, we sequenced mouse DNA related to human chromosome 19. Comparative sequence alignments yielded confirmatory evidence for hypothetical genes and identified exons, regulatory elements, and candidate genes that were missed by other predictive methods. Chromosome-wide comparisons revealed a difference between single-copy HSA19 genes, which are overwhelmingly conserved in mouse, and genes residing in tandem familial clusters, which differ extensively in number, coding capacity, and organization between the two species. Finally, we sequenced breakpoints of all 15 evolutionary rearrangements, providing a view of the forces that drive chromosome evolution in mammals.

Continuous monitoring of the environment for infectious diseases and related biowarfare agents requires the implementation of practical cost-effective methodologies that are highly sensitive and specific. One compatible method employed in clinical diagnostics is real-time polymerase chain reaction (PCR) analysis. The utility of this technique for environmental monitoring is limited, however, by the utilization of single-use consumables in commercial PCR instruments. This greatly increases mechanical complexity, because sophisticated robotic mechanisms must replenish the disposable elements. An alternative strategy develops an autonomous monitoring system consisting of reusable modules that readily interface with fluidic circuitry in a flow-through scheme. The reduced complexity should increase reliability while decreasing operating costs. In this report, we describe a reusable, flow-through PCR module that functions as one component in such a system. This module was rigorously evaluated with Bacillus anthracis genomic DNA and demonstrated high repeatability, sensitivity, and efficiency, with no evidence of sample-to-sample carryover.

Based on 2D 'H-IH and 2D and 3D 'H-"N NMR spectroscopies, complete ' H NMR assignments are reported for zinc-containing Clostridium pasteurianum rubredoxin (Cp ZnRd). Complete 'H NMR assignments are also reported for a mutated Cp ZnRd, in which residues near the N-terminus, namely, Met 1 , Lys 2, and Pro 15, have been changed to their counterparts, (-), Ala and Glu, respectively, in rubredoxin from the hyperthermophilic archaeon, Pyrococcusfuriosus (Pf Rd). The secondary structure of both wild-type and mutated Cp ZnRds, as determined by NMR methods, is essentially the same. However, the NMR data indicate an extension of the threestranded &sheet in the mutated Cp ZnRd to include the N-terminal Ala residue and Glu 15, as occurs in Pf Rd. The mutated Cp Rd also shows more intense NOE cross peaks, indicating stronger interactions between the strands of the 0-sheet and, in fact, throughout the mutated Rd. However, these stronger interactions do not lead to any significant increase in thermostability, and both the mutated and wild-type Cp Rds are much less thermostable than Pf Rd. These correlations strongly suggest that, contrary to a previous proposal [Blake PR et al., 1992, Protein Sci I : 1508-15211, the thermostabilization mechanism of Pf Rd is not dominated by a unique set of hydrogen bonds or electrostatic interactions involving the N-terminal strand of the 0-sheet. The NMR results also suggest that an overall tighter protein structure does not necessarily lead to increased thermostability.Keywords: Clostridium pasteaurianum; 2D NMR; 3D NMR; mutated rubredoxin; protein secondary structure; protein thermostability; rubredoxin; zinc-containing rubredoxin The group of bacterial and archaeal iron-sulfur proteins known as rubredoxins appears to be ideal for delineating molecular determinants of non-heme metalloprotein structure, thermostability, and redox properties. Rds have in common the following favorable characteristics for such investigations: small size (typ- ically 52-54 residues), a very simple redox active Fe(SCys), site (whose Cys residues are underlined in Scheme I), a large and continually expanding database of amino acid sequences (Zeng et al., 1995), at least five high-resolution X-ray crystal structures (Sieker et al., 1994), a characteristic structural motif featuring a three-stranded @-sheet, and well-characterized proteins from several mesophiles and from the hyperthermophilic archaeon Pyrococcus furiosus. Pf Rd is known to be remarkably thermostable compared to Rds from mesophilic organisms (Blake et al., 1991;Day et al., 1992). However, there are no obvious distinguishing structural or spectroscopic features of the FeS, site in Pf Rd compared to Rds from mesophiles. Instead, based on the NMR solution structure of the zinc-substituted Pf Rd (Blake et al., 1992) and the X-ray crystal structure of Pf FeRd (Day et al., 1992), a unique set of hydrogen bonds and electrostatic interactions was proposed to inhibit "unzipping" of the threestranded @-sheet at high temperatures. This inhibition cou...