Two-dimensional gel electrophoresis technique for determination of the crosslinked nucleotides in cleavable covalent RNA-RNA complexes. Application to Escherichia coli and Bacillus subtilis acetylvalyl-tRNA covalently linked to E. coli 16 S and yeast 18 S ribosomal RNA

Biochemistry ◽  
1984 ◽  
Vol 23 (3) ◽  
pp. 438-445 ◽  
Author(s):  
Chantal Ehresmann ◽  
James Ofengand
Keyword(s):  
Ribosomal Rna ◽  
Gel Electrophoresis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
E Coli ◽  
Covalently Linked ◽  
Rna Complexes ◽  
Electrophoresis Technique ◽  
18 S Ribosomal Rna

scholarly journals Analysis of H-2 and Ia molecules by two-dimensional gel electrophoresis.

1977 ◽  
Vol 146 (5) ◽  
pp. 1261-1279 ◽  
Author(s):  
P P Jones
Keyword(s):  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Surface Proteins ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Cell Surface Proteins ◽  
Ia Antigens ◽  
Polypeptide Chains ◽  
Kinetics Of ◽  
Electrophoresis Technique

Mouse lymphocyte H-2 and Ia glycoproteins have been analyzed with a two-dimensional (2-D) acrylamide gel electrophoresis technique, in which proteins are separated first according to their charge in isoelectrofocusing gels and then according to their size in sodium dodecyl sulfate gels. Individual polypeptide chains from radiolabeled cells are resolved as discrete spots on autoradiograms of the gels, forming patterns which are characteristic of the proteins in the sample. 2-D gels of H-2K, H-2D, and Ia glycoproteins immunoprecipitated from 35S-methionine-labeled cells reveal that these proteins exist in the cells as complex arrays of molecules heterogeneous in both size and charge. Lactoperoxidase-catalyzed radioiodination of lymphocyte surfaces labels only subsets of the total H-2 and Ia molecules with 125I, indicating that some of the molecules may represent cytoplasmic precursors of the cell surface proteins. This theory is supported by the kinetics of labeling of various spots in 35S-methionine pulse-chase experiments. The 2-D gel patterns obtained for both H-2 and Ia antigens have also been shown to be haplotype-specific and independent of the genetic background.

scholarly journals IDENTIFICATION OF THE BACTERIOPHAGE T4 unf (=alc) GENE PRODUCT, A PROTEIN INVOLVED IN THE SHUTOFF OF HOST TRANSCRIPTION

Genetics ◽  
1984 ◽  
Vol 108 (2) ◽  
pp. 305-317
Author(s):  
Richard E Herman ◽  
Nancy Haas ◽  
D Peter Snustad
Keyword(s):  
Gene Product ◽  
Gel Electrophoresis ◽  
Gene Dosage ◽  
Bacteriophage T4 ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
E Coli ◽  
Phage T4 ◽  
Protein Functions

ABSTRACT The introduction of plasmid pR386 into E. coli cells renders them restrictive to the growth of phage T4 unf (=alc) mutants. This system has been used to isolate Unf+ revertants, which, along with the mutant parental strains, have been used to identify the unf gene product by two-dimensional gel electrophoresis. Synthesis of the unf gene product, a polypeptide of just over 18,000 daltons in size, begins within 1 min after infection and terminates at about 12 min after infection at 30°. Gene dosage experiments suggest that the unf protein functions catalytically.

scholarly journals CspI, the Ninth Member of the CspA Family ofEscherichia coli, Is Induced upon Cold Shock

1999 ◽  
Vol 181 (5) ◽  
pp. 1603-1609 ◽  
Author(s):  
Nan Wang ◽  
Kunitoshi Yamanaka ◽  
Masayori Inouye
Keyword(s):  
Gel Electrophoresis ◽  
Untranslated Region ◽  
Cold Shock ◽  
Two Dimensional ◽  
Optimal Temperature ◽  
Two Dimensional Gel Electrophoresis ◽  
E Coli ◽  
Temperature Range ◽  
Negative Effect ◽  
Temperature Ranges

ABSTRACT Escherichia coli contains the CspA family, consisting of nine proteins (CspA to CspI), in which CspA, CspB, and CspG have been shown to be cold shock inducible and CspD has been shown to be stationary-phase inducible. The cspI gene is located at 35.2 min on the E. coli chromosome map, and CspI shows 70, 70, and 79% identity to CspA, CspB, and CspG, respectively. Analyses of cspI-lacZ fusion constructs and thecspI mRNA revealed that cspI is cold shock inducible. The 5′-untranslated region of the cspI mRNA consists of 145 bases and causes a negative effect on cspIexpression at 37°C. The cspI mRNA was very unstable at 37°C but was stabilized upon cold shock. Analyses of the CspI protein on two-dimensional gel electrophoresis revealed that CspI production is maximal at or below 15°C. Taking these results together, E. coli possesses a total of four cold shock-inducible proteins in the CspA family. Interestingly, the optimal temperature ranges for their induction are different: CspA induction occurs over the broadest temperature range (30 to 10°C), CspI induction occurs over the narrowest and lowest temperature range (15 to 10°C), and CspB and CspG occurs at temperatures between the above extremes (20 to 10°C).

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