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R Staden

Publications and source records attributed to R Staden.

At least 55 records · Page 3Linked to original sources

Software for genome mapping by fingerprinting techniques.

A genome mapping package has been developed for reading and assembling data from clones analysed by restriction enzyme fragmentation and polyacrylamide gel electrophoresis. The package comprises: data entry; matching; assembly; statistical analysis; modelling. Data entry can be either manual or by a semiautomatic system based on a scanning densitometer. The primary emphasis in the analytical routines is on flexibility and interactive convenience, so that the operator has full knowledge of and control over the growing map, but a variety of automatic options are included. The package continually grows to meet the needs of the Caenorhabditis project.

Base Sequence↗

Methods to define and locate patterns of motifs in sequences.

A method to define and search for complex patterns of motifs in nucleic acid and protein sequences is described. With this method nucleic acid motifs can be defined in eight different ways and protein motifs in six. A pattern is defined by a list of motifs. The motifs in a list are combined using the logical operators AND, OR and NOT. The list also defines the ranges of allowed separations of the motifs in the pattern. Programs to search for patterns in individual sequences and libraries of sequences are described. Patterns are defined by users and stored as annotated disk files. Hence the programming to define and locate new structures can be performed by users and fewer specific novel algorithms should be required. Examples are given of searches for transcription initiation regions, nematode mitochondrial tRNA genes and for members of the globin sequence family.

Algorithms↗

The current status and portability of our sequence handling software.

I describe the current status of our sequence analysis software. The package contains a comprehensive suite of programs for managing large shotgun sequencing projects, a program containing 61 functions for analysing single sequences and a program for comparing pairs of sequences for similarity. The programs that have been described before have been improved by the addition of new functions and by being made very much easier to use. The major interactive programs have 125 pages of online help available from within them. Several new programs are described including screen editing of aligned gel readings for shotgun sequencing projects; a method to highlight errors in aligned gel readings, new methods for searching for putative signals in sequences. We use the programs on a VAX computer but the whole package has been rewritten to make it easy to transport it to other machines. I believe the programs will now run on any machine with a FORTRAN77 compiler and sufficient memory. We are currently putting the programs onto an IBM PC XT/AT and another micro running under UNIX.

Amino Acid Sequence↗

A method for measuring the non-random bias of a codon usage table.

We describe a new statistical method for measuring bias in the codon usage table of a gene. The test is based on the multinomial and Poisson distributions. The method is used to scan DNA sequences and measure the strength of codon preference. For E. Coli we show that the strength of codon preference is related to levels of gene expression. The method can also be used to compare base triplet frequencies with those expected from the base composition. This second type of codon bias test is useful for distinguishing coding from non-coding regions.

Base Sequence↗

Computer methods to locate signals in nucleic acid sequences.

This paper describes computer methods for locating signals in nucleic acid sequences. The signals include ribosome binding sites, promoter sequences and splice junctions. The methods are of use both to those trying to interpret the function of newly determined sequences and to those studying the molecular mechanisms involved in the recognition of these special signal sequences.

Base Sequence↗

Graphic methods to determine the function of nucleic acid sequences.

We describe an interactive computer program (ANALYSEQ) that is used from a simple graphics terminal. The main purpose of the program is to determine the function of nucleic acid sequences but it also offers the simpler listing, searching and counting options. It contains methods to locate genes by looking for the effects that coding for a protein has on the coding sequence, to locate tRNA genes by looking for secondary structure and conserved bases, and methods to locate signals such as promoters. Techniques to identify unusual regions of sequence and to search for potential Z DNA-forming regions are also included. Most of the routines produce graphical output which gives ease of interpretation and allows superposition of several independent forms of analysis.

Animals↗

Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing.

This paper describes a computer method for handling gel reading data produced by the shotgun method of DNA sequencing. The method greatly reduces the time the sequencer needs to spend checking and editing his data and yet it produces a consensus sequence for which the accuracy of determination of every base can be clearly shown. The program can take a batch of new gel readings, screen them against vector sequences removing any that match, and then compare and align all the sequences to produce a final consensus. No information is lost in this process as alignments are achieved by making only insertions and because all the individual gel readings are added to a database from which they can be retrieved and displayed lined up one above the other. This allows the user to check on the alignments achieved by the program and if necessary change them. As each gel reading is added to the database the consensus is automatically updated accordingly and used for the next comparisons. This is a much faster process than comparing each new gel against every individual gel in the database.

Autoanalysis↗

An interactive graphics program for comparing and aligning nucleic acid and amino acid sequences.

This paper describes a computer program designed to look for similarities between pairs of nucleic or amino acid sequences. The program looks both for segments of perfect identity or for regions where, using a scoring matrix, a minimum value is exceeded. The results of comparisons are presented as a matrix which is displayed on a simple graphics terminal. Use of a graphics terminal allows the user to display the whole of the two sequences in one screenful or to home-in on regions of interest to examine them in more detail. The program is interactive and so the user can easily see the effect of changes to variables and can use inbuilt editing functions to make insertions to produce alignments of the two sequences. These aligned sequences can then be saved on disk files for further processing.

Amino Acid Sequence↗

Complete nucleotide sequence of the chicken chromosomal ovalbumin gene and its biological significance.

The nucleotide sequence of the entire chicken chromosomal ovalbumin gene has been determined. The gene is 7564 nucleotides in length to code for a mature messenger RNA of 1872 nucleotides. Comparison of the sequence at the 5'-terminal region of the gene with that reported by others has revealed multiple polymorphic nucleotides in the structural, intervening, and flanking DNA sequences. Some of the polymorphic sites occur at positions very close to splice junctions or the eucaryotic promoter sequence, yet apparently have little or no effect on the expression of this gene. The heptanucleotide promoter sequence TATATAT present in the 5'-flanking region of the ovalbumin gene does not occur within the confines of the gene. Nevertheless, multiple Hogness box sequences similar to those found in other eucaryotic genes were delineated within the boundaries of the gene. These internal Hogness box sequences are not used for transcription initiation. Similarly, the hexanucleotide sequence AATAAA common to all eucaryotic messenger RNAs at the 3'-untranslated region occurs seven additional times within the ovalbumin gene. These sites are not used for transcription termination or polyadenylation. Thus, although these sequences may play important roles in the initiation or termination of gene transcripts as well as polyadenylation of the transcripts, the specificity for such biological functions must not reside within these sequences alone. Furthermore, sequences complementary to the highly conserved rat U1 small nuclear RNA have been found throughout the gene. Many of these regions of complementarity occur in the structural sequences. If the small nuclear RNA does play a role in splicing, the specificity must be provided also by other as yet undefined components.

Animals↗

Sequence and organization of the human mitochondrial genome.

The complete sequence of the 16,569-base pair human mitochondrial genome is presented. The genes for the 12S and 16S rRNAs, 22 tRNAs, cytochrome c oxidase subunits I, II and III, ATPase subunit 6, cytochrome b and eight other predicted protein coding genes have been located. The sequence shows extreme economy in that the genes have none or only a few noncoding bases between them, and in many cases the termination codons are not coded in the DNA but are created post-transcriptionally by polyadenylation of the mRNAs.

Base Sequence↗

A new computer method for the storage and manipulation of DNA gel reading data.

This paper describes a new way of storing DNA gel reading data and an accompanying set of computer programs. These programs will perform all the manipulations that are required on data gained by the so-called 'shotgun' method of DNA sequencing. This system simplifies the computer processing involved with this sequencing method and also has the capability of being able at any time during a project to display, lined up in register, all the gel reading covering any section of the sequence.

Base Sequence↗

Different pattern of codon recognition by mammalian mitochondrial tRNAs.

Analysis of an almost complete mammalian mitochondrial DNA sequence has identified 23 possible tRNA genes and we speculate here that these are sufficient to translate all the codons of the mitochondrial genetic code. This number is much smaller than the minimum of 31 required by the wobble hypothesis. For each of the eight genetic code boxes with four codons for one amino acid we find a single specific tRNA gene with T in the first (wobble) position of the anticodon. We suggest that these tRNAs with U in the wobble position can recognize all four codons in these genetic code boxes either by a "two out of three" base interaction or by U.N wobble.

Animals↗

A strategy of DNA sequencing employing computer programs.

With modern fast sequencing techniques and suitable computer programs it is now possible to sequence whole genomes without the need of restriction maps. This paper describes computer programs that can be used to order both sequence gel readings and clones. A method of coding for uncertainties in gel readings is described. These programs are available on request.

Base Sequence↗