scholarly journals Polymerase Chain Reaction and Restriction Endonuclease Digestion for Selected Members of the “Mycoplasma Mycoides Cluster” and Mycoplasma Putrefaciens

1997 ◽  
Vol 9 (2) ◽  
pp. 186-190 ◽  
Author(s):  
Jose L. Rodriguez ◽  
Richard W. Ermel ◽  
Thomas P. Kenny ◽  
Dale L. Brooks ◽  
Al J. DaMassa
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Endonuclease ◽  
Restriction Enzymes ◽  
The United States ◽  
Restriction Endonuclease Digestion ◽  
Chain Reaction ◽  
Large Colony ◽  
Mycoplasma Mycoides ◽  
Polymerase Chain ◽  
Endonuclease Digestion

A specific diagnostic method using the polymerase chain reaction, together with restriction endonuclease digestion patterns, was developed for members of the “ Mycoplasma mycoides cluster” that normally occur in the United States (i.e., Mycoplasma mycoides subsp. mycoides Large Colony and Mycoplasma capricolum subsp. capricolum in addition to “cluster” mycoplasma, bovine serogroup 7, and Mycoplasma putrefaciens. The digestion of “cluster” polymerase chain reaction DNA (1,225 bp) amplification products with restriction enzymes Asel and SspI gave mycoplasma species-specific patterns for all strains of M. mycoides subsp. mycoides Large Colony, M. capricolum subsp. capricolum, and bovine group 7 tested. Moreover, we found a nonspecific amplification product for M. putrefaciens that occurred with the oligonucleotide primers used for the “ M. mycoides cluster” reaction. However, the restriction endonuclease digestion patterns observed with the restriction enzymes AluI, AseI, and SspI for M. putrefaciens were different than the digestion patterns obtained for the other “cluster” mycoplasmas. This report confirms the usefulness of polymerase chain reaction DNA amplification allied with restriction enzyme digestion profile analysis for the rapid and specific identification of mycoplasmas belonging to the “ M. mycoides cluster” and M. putrefaciens.

scholarly journals Human fibrinogen polymorphic site analysis by restriction endonuclease digestion and allele-specific polymerase chain reaction amplification: identification of polymorphisms at positions A alpha 312 and B beta 448

Blood ◽  
1993 ◽  
Vol 82 (7) ◽  
pp. 2117-2124 ◽  
Author(s):  
RE Baumann ◽  
AH Henschen
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Endonuclease ◽  
Dna Sequence ◽  
Restriction Analysis ◽  
Restriction Endonuclease Digestion ◽  
Chain Reaction ◽  
Specific Polymerase Chain Reaction ◽  
Allele Specific ◽  
Polymerase Chain ◽  
Endonuclease Digestion

Abstract In the fibrinogen molecule, a total of seven sites have been tentatively identified as polymorphic; however, disagreements about these sites have been observed among the various protein and DNA sequence data published. To allow examination of the potential polymorphic sites at the DNA level, human genomic DNA samples were prepared from 110 unrelated, healthy individuals. Either allele- specific polymerase chain reaction (ASPCR) amplification or PCR amplification followed by restriction endonuclease digestion was used to detect the presence of possible polymorphisms. Two polymorphic sites were confirmed, one at A alpha 312 (Thr/Ala) by RsaI restriction analysis, and a second at B beta 448 (Arg/Lys) by MnlI restriction analysis. Mendelian inheritance of both polymorphisms was demonstrated and allele frequencies were estimated as 0.76/0.24 and 0.85/0.15 for the A alpha 312 and B beta 448 sites, respectively. The sites at A alpha 47, A alpha 296, B beta 162, B beta 296, and gamma 88 showed no evidence of variation in any of our samples. The amino acid polymorphisms at A alpha 312 and B beta 448 reflect conservative residue changes with unknown effects on fibrinogen structure or function. An additional, previously unrecognized DNA sequence variant was detected in a single individual in the second intron of the A alpha chain using HinfI restriction analysis.

scholarly journals Human fibrinogen polymorphic site analysis by restriction endonuclease digestion and allele-specific polymerase chain reaction amplification: identification of polymorphisms at positions A alpha 312 and B beta 448

Blood ◽  
1993 ◽  
Vol 82 (7) ◽  
pp. 2117-2124 ◽  
Author(s):  
RE Baumann ◽  
AH Henschen
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Endonuclease ◽  
Dna Sequence ◽  
Restriction Analysis ◽  
Restriction Endonuclease Digestion ◽  
Chain Reaction ◽  
Specific Polymerase Chain Reaction ◽  
Allele Specific ◽  
Polymerase Chain ◽  
Endonuclease Digestion

In the fibrinogen molecule, a total of seven sites have been tentatively identified as polymorphic; however, disagreements about these sites have been observed among the various protein and DNA sequence data published. To allow examination of the potential polymorphic sites at the DNA level, human genomic DNA samples were prepared from 110 unrelated, healthy individuals. Either allele- specific polymerase chain reaction (ASPCR) amplification or PCR amplification followed by restriction endonuclease digestion was used to detect the presence of possible polymorphisms. Two polymorphic sites were confirmed, one at A alpha 312 (Thr/Ala) by RsaI restriction analysis, and a second at B beta 448 (Arg/Lys) by MnlI restriction analysis. Mendelian inheritance of both polymorphisms was demonstrated and allele frequencies were estimated as 0.76/0.24 and 0.85/0.15 for the A alpha 312 and B beta 448 sites, respectively. The sites at A alpha 47, A alpha 296, B beta 162, B beta 296, and gamma 88 showed no evidence of variation in any of our samples. The amino acid polymorphisms at A alpha 312 and B beta 448 reflect conservative residue changes with unknown effects on fibrinogen structure or function. An additional, previously unrecognized DNA sequence variant was detected in a single individual in the second intron of the A alpha chain using HinfI restriction analysis.

scholarly journals Transmission of the Citrus Variegated Chlorosis Bacterium Xylella fastidiosa with the Sharpshooter Oncometopia nigricans

Plant Disease ◽  
2002 ◽  
Vol 86 (11) ◽  
pp. 1237-1239 ◽  
Author(s):  
R. H. Brlansky ◽  
V. D. Damsteegt ◽  
J. S. Hartung
Keyword(s):  
United States ◽  
Polymerase Chain Reaction ◽  
Xylella Fastidiosa ◽  
The United States ◽  
Reliable Indicator ◽  
Symptom Development ◽  
Chain Reaction ◽  
Citrus Variegated Chlorosis ◽  
Specific Polymerase Chain Reaction ◽  
Polymerase Chain

Citrus variegated chlorosis (CVC) is an economically important, destructive disease in Brazil and is caused by the bacterium Xylella fastidiosa Wells. The bacterium has been found to be transmitted in Brazil by sharpshooter leafhoppers (Cicadellidae). Sharpshooters are present in most citrus growing areas of the United States. The sharpshooter leafhopper, Oncometopia nigricans Walker, frequently is found feeding on citrus in Florida. This sharpshooter transmits the X. fastidiosa strains that cause Pierce's disease of grape and ragweed stunt. Research was initiated to determine if O. nigricans was capable of vectoring the X. fastidiosa that causes CVC. In 59 different transmission tests, using 1 to 57 insects per test, transmission of the bacterium was observed 12 times (20.3%). Symptom development in the greenhouse was not a reliable indicator of transmission. Transmission was verified by specific polymerase chain reaction-based assays. Individual insects were able to transmit the bacterium. This information on sharpshooter transmission of CVC is needed to assess the threat posed by the CVC disease to the citrus industries in the United States.

Sign in / Sign up

Export Citation Format

Share Document