Biomedical subjects

T D McKnight

Publications and source records attributed to T D McKnight.

22 records · Page 2Linked to original sources

PubMed1986

Nucleotide sequence and molecular evolution of two tomato genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase.

We have isolated and sequenced two cDNA clones (LESS5 and LESS17) encoding the small subunit of ribulose-1,5-bisphosphate carboxylase of tomato (Lycopersicon esculentum). At the nucleotide level, the protein-coding regions of these genes are 85% conserved, while the untranslated 3' regions are only 55% conserved. Comparison with rbcS genes from other species of Solanaceae suggests that the tomato LESS5 gene, the Nicotiana tabacum NTSS23 gene and the Petunia hybrida SSU8 gene are orthologous members of the rbcS gene family. In addition, the tomato gene LESS17, and the Petunia hybrida gene SSU611, may also be orthologous, since their untranslated 3' regions are related. There is a large difference between the two tomato rbcS genes in the frequency of the CG dinucleotide. This difference may reflect different levels of methylation, and therefore expression, of the tomato genes. Many of the differences involving the CG dinucleotide can be represented as transitions between C and T on the noncoding strand. Such changes are consistent with observations that methylated cytosines are hot-spots for transitions.

Base Sequence↗

PubMed1984

A simplified and efficient vector-primer cDNA cloning system.

A simplified, efficient, and versatile vector-primer cDNA cloning system is presented. The dimer-primer system is a modification of the method of Okayama and Berg (1982) with the following features: (i) the vector-primer molecules are more rapidly and reliably prepared by virtue of the elimination of an endonuclease digestion and the agarose gel purification step from the original method, and (ii) the final cDNA products contain polylinkers at both cDNA-vector junctions, simplifying the size analysis, subcloning, and sequencing of inserts. The system is highly efficient, yielding greater than 10(5) transformants using 1 microgram mRNA and 1 pmol of vector-primer ends, with 75% or more of the transformants having inserts. The ability of the system to produce clones of full-length or near full-length is demonstrated by the analysis of 32 ribulose-1,5-bisphosphate carboxylase small subunit cDNA clones from tomato.

Animals↗

PubMed1983

Characterization and preliminary mapping of cauliflower mosaic virus transcripts.

Using Northern blot analysis we have studied the transcription of the CM4-184 Ga. strain of cauliflower mosaic virus (CaMV) DNA. This analysis reveals that this CaMV strain, like the Cabb-BS and the Cabb B-JI strains, produces both a genomic length transcript and a 1900-nucleotide (nt) transcript during infection. In addition, we detect an 1800-nt PA+ transcript mapping primarily to the EcoRI-c region of the virus, and three apparent minor viral-specific PA + RNAs of 4900, 4500, and 4300 nt. We also report the presence of two small viral single-stranded DNAs produced during infection, and show that both DNAs are derived from the EcoRI-b region of the virus.

Chromosome Mapping↗

PubMed1981

Isolation and Mapping of Small Cauliflower Mosaic Virus DNA Fragments Active as Promoters in Escherichia coli.

Small EcoRI(*) fragments of cauliflower mosiac virus DNA (strain CM4-184), which act as promoters for the tetracycline resistance gene on the promoter probe plasmid pBRH4 in Escherichia coli, have been isolated and mapped on the viral genome. Two regions of the viral genome contain DNA sequences with promoter activity in E. coli. Two independent cloned fragments from one region direct a high level of tetracycline resistance (up to 38 mug of tetracycline per ml). Two independent fragments from the second region of the viral genome also direct tetracycline resistance, but at lower levels. The activity of the two fragments with the strongest promoter activity in E. coli may direct transcription of the viral genome in a clockwise direction. This is consistent with the direction of transcription predicted from sequence analysis of the viral DNA (Franck et al., Cell 21: 285-294, 1980). One of these fragments maps at the start of a large open translational reading frame which is predicted to contain the coding sequence for the viral coat protein. Each promoter-active fragment is located in the 5'-terminal portion of one of the six open reading frames predicted from the DNA sequence.

Journal Article↗