Objective: To study the relationship between the polymorphism of drug resistant gene rpoB and drug resistance against rifampicin(RFP) of M. tuberculosis L-forms, and to evaluate its clinical application. Methods: A to...Objective: To study the relationship between the polymorphism of drug resistant gene rpoB and drug resistance against rifampicin(RFP) of M. tuberculosis L-forms, and to evaluate its clinical application. Methods: A total of 52 clinical isolated strains of M. tuberculosis L-forms were collected. rpoB gene polymorphism was analyzed by polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) and conventional antimicrobial susceptibility test (AST). Their results were compared. Results: AST results showed that 38 of 52 clinical isolated strains were drug resistance (73.08％),while PCR-SSCP indicated 65.38％ (32/52) rpoB gene polymorphism. There was no statistic significance(χ2= 2.4914) between the 2 methods. Conclusion:Combined the application of PCR-SSCP with AST in detecting rpoB drug resistant gene polymorphism of M. tuberculosis L-form from pneumoconiosis patients with tuberculosis may have advantages at earlier diagnosis and guidance of clinical medications.展开更多
By employing different gel components and electrophoresis conditions, the PCR-SSCP(polymerase chain reaction-single-stranded conformation polymorphism) method was used to detect the polymorphism(CCC or CGC ) of codon ...By employing different gel components and electrophoresis conditions, the PCR-SSCP(polymerase chain reaction-single-stranded conformation polymorphism) method was used to detect the polymorphism(CCC or CGC ) of codon 72 in exon 4 of p53 gene. The results展开更多
We reported our preliminary study on mapping of the expressed sequence tag (EST) using single-stranded DNA conformation polymorphism (SSCP) on polymerase chain reaction (PCR) product. Five ESTs primer pairs, developed...We reported our preliminary study on mapping of the expressed sequence tag (EST) using single-stranded DNA conformation polymorphism (SSCP) on polymerase chain reaction (PCR) product. Five ESTs primer pairs, developed from nuclear genes including CAD(cinnamyl-alcohol dehydrgenase), CHS(chalcone synthase), NIR(nitrite reductase), ARA554 (cDNA expressed in differentiating xylem in Pinus taedae) and GS(glutamine synthetase), were selected to optimize the experimental protocol and generate mapping markers in the megagemtophytes from a single tree of Pinus massoniana. Our ultimate goal was to use SSCP approach to construct a transcriptional map for comparative mapping studies in P. massoniana. The efficiency to construct a transcriptional map in P. massoniana based on the published EST primer pairs derived from other pine species critically depended on the successful amplification of EST fragments and the ratio of the heterozygous loci revealed in P.massoniana. In this study, 3 (60) out of 5 tested EST primer pairs were succeeded in amplification, however, only 1(20) gene was heterozygous in the tested tree. The ratio of heterozygous loci detected in this study was similar to that revealed by anonymous marker in P.taeda. Therefore, a low efficiency would be expected if the map would be constructed using single pedigree. The segregation ratio of loci revealed by primer pairs would be higher if multiple pedigrees would be used. We proposed that consensus mapping approach based on multiple-pedigree should be used for EST mapping and therefore to increase the efficiency of EST mapping.展开更多
Objective：To study the genetic variation of mitochondrial DNA （mtDNA） among common laboratory strains of inbred mice. Methods： The genetic polymorphism of mtDNA among 4 classical laboratory strains of inbred mice ...Objective：To study the genetic variation of mitochondrial DNA （mtDNA） among common laboratory strains of inbred mice. Methods： The genetic polymorphism of mtDNA among 4 classical laboratory strains of inbred mice and 3 inbred strains of mice established in China was analyzed by polymerase chain reaction coupled with restriction fragment length polymorphism（PCR-RFLP） and PCR coupled with single-stranded conformation polymorphism（PCR-SSCP）. Results： With regard to the D-loop （Displacement loop, D-loop）, tRNA^ Met＋Glu＋Ile, and ND3 （NADH dehydrogenase subunit 3, ND3） gene fragments of mtDNA from these mice,no variation was revealed by PCR-RFLP at 46 restriction enzyme sites. Further analyzed by PCR-SSCP,the D-loop 5＇fragment and 3＇end fragment of mtDNA from these mice also showed no genetic variation. Conclusion： Owing to maternal mode of inheritance of mtDNA,the results indicate that these common inbred strains of mice share the same maternal lineage.展开更多
文摘Objective: To study the relationship between the polymorphism of drug resistant gene rpoB and drug resistance against rifampicin(RFP) of M. tuberculosis L-forms, and to evaluate its clinical application. Methods: A total of 52 clinical isolated strains of M. tuberculosis L-forms were collected. rpoB gene polymorphism was analyzed by polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) and conventional antimicrobial susceptibility test (AST). Their results were compared. Results: AST results showed that 38 of 52 clinical isolated strains were drug resistance (73.08％),while PCR-SSCP indicated 65.38％ (32/52) rpoB gene polymorphism. There was no statistic significance(χ2= 2.4914) between the 2 methods. Conclusion:Combined the application of PCR-SSCP with AST in detecting rpoB drug resistant gene polymorphism of M. tuberculosis L-form from pneumoconiosis patients with tuberculosis may have advantages at earlier diagnosis and guidance of clinical medications.
文摘By employing different gel components and electrophoresis conditions, the PCR-SSCP(polymerase chain reaction-single-stranded conformation polymorphism) method was used to detect the polymorphism(CCC or CGC ) of codon 72 in exon 4 of p53 gene. The results
文摘We reported our preliminary study on mapping of the expressed sequence tag (EST) using single-stranded DNA conformation polymorphism (SSCP) on polymerase chain reaction (PCR) product. Five ESTs primer pairs, developed from nuclear genes including CAD(cinnamyl-alcohol dehydrgenase), CHS(chalcone synthase), NIR(nitrite reductase), ARA554 (cDNA expressed in differentiating xylem in Pinus taedae) and GS(glutamine synthetase), were selected to optimize the experimental protocol and generate mapping markers in the megagemtophytes from a single tree of Pinus massoniana. Our ultimate goal was to use SSCP approach to construct a transcriptional map for comparative mapping studies in P. massoniana. The efficiency to construct a transcriptional map in P. massoniana based on the published EST primer pairs derived from other pine species critically depended on the successful amplification of EST fragments and the ratio of the heterozygous loci revealed in P.massoniana. In this study, 3 (60) out of 5 tested EST primer pairs were succeeded in amplification, however, only 1(20) gene was heterozygous in the tested tree. The ratio of heterozygous loci detected in this study was similar to that revealed by anonymous marker in P.taeda. Therefore, a low efficiency would be expected if the map would be constructed using single pedigree. The segregation ratio of loci revealed by primer pairs would be higher if multiple pedigrees would be used. We proposed that consensus mapping approach based on multiple-pedigree should be used for EST mapping and therefore to increase the efficiency of EST mapping.
基金Supported by Developmental Programming Item of National Keystone Basic Research (G2000016106) and National Natural Science Foundation of China (No. 39600079)
文摘Objective：To study the genetic variation of mitochondrial DNA （mtDNA） among common laboratory strains of inbred mice. Methods： The genetic polymorphism of mtDNA among 4 classical laboratory strains of inbred mice and 3 inbred strains of mice established in China was analyzed by polymerase chain reaction coupled with restriction fragment length polymorphism（PCR-RFLP） and PCR coupled with single-stranded conformation polymorphism（PCR-SSCP）. Results： With regard to the D-loop （Displacement loop, D-loop）, tRNA^ Met＋Glu＋Ile, and ND3 （NADH dehydrogenase subunit 3, ND3） gene fragments of mtDNA from these mice,no variation was revealed by PCR-RFLP at 46 restriction enzyme sites. Further analyzed by PCR-SSCP,the D-loop 5＇fragment and 3＇end fragment of mtDNA from these mice also showed no genetic variation. Conclusion： Owing to maternal mode of inheritance of mtDNA,the results indicate that these common inbred strains of mice share the same maternal lineage.
