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Biomedical subjects

Y X Pan

Publications and source records attributed to Y X Pan.

44 records · Page 3Linked to original sources
PubMed1994

[Specific detection of Mycobacterium tuberculosis in clinical material by PCR and Southern blot].

An amplified fragment of 245 base pairs from IS-986 sequence hybridized specifically with DNA of M. tuberculosis complex strains. The fragment was labelled with digoxigenin. After amplification by PCR and southern blot hybridization with the dig-labelled probe, the limit of detection of purified genomic M. tuberculosis DNA amounted to 1 fg. We detected sputum, pleural and synovial fluid with active tuberculosis by PCR and southern blot, and the sensitivities were 81.0%, 64.2% and 57.7%, respectively. From the result of the experiment, it is a very useful method for diagnosis of tuberculosis by combining PCR with Southern blot.

Blotting, Southern↗
PubMed1993

Saccharomyces cerevisiae expression of exogenous vacuolar ATPase subunits B.

The precise function of subunit B of the vacuolar H(+)-ATPase class is unknown, but it is essential for proton pumping. We have previously reported the DNA sequence and predicted protein sequence of the vacuolar ATPase subunit B for Candida tropicalis (Gu, H.H., Gallagher, M.J., Rupkey, S. and Dean, G.E. (1990) Nucleic Acids Res. 18, 7446). When the Candida gene was expressed in a Saccharomyce cerevisiae delta vat2 mutant from which the homologous gene had been deleted, viability and vacuolar acidification was restored to apparently wild-type levels. The predicted identity between these two proteins is 90%. We have searched for vacuolar ATPase subunits B from other species that might show a difference in function, when expressed in yeast, relative to the endogenous gene. We have cloned an apparently full-length 1.8-kb bovine subunit B cDNA from adrenal medulla that is about 1 kb shorter than the previously reported bovine brain cDNA (Puopolo, K., Kumamoto, C., Adachi, I., Magner, R. and Forgac, M. (1992) J. Biol. Chem. 267, 3696-3706; Nelson, R.D., Guo, X.L., Masood, K., Brown, D., Kalkbrenner, M. and Gluck, S. (1992) Proc. Natl. Acad. Sci. USA 89, 3541-3545), but nearly identical throughout the coding nucleotide and protein sequences; it is only 74% identical to the Saccharomyces subunit B protein sequence. Upon expression of this cDNA in two different delta vat2 deletion strains, the bovine cDNA restored function only partially, as judged by both viability at high pH and vacuolar acidification. Current work is aimed at determining which regions of the bovine protein require alteration in order to fully restore the delta vat2 strain to wild-type acidification, with the eventual goal of identifying interactive residues between subunit B and other proteins required for pump function.

Amino Acid Sequence↗
PubMed1993

[A study on the diagnosis of pulmonary tuberculosis and silicotuberculosis by PCR].

The sputa of 156 patients with lung disease were detected by smear, culture and PCR for mycobacterium tuberculosis. The detectable limitation was 10 fg DNA and 5 organisms. Both sensitivity and specificity of detection by PCR were much better than smear or culture techniques. Our results demonstrated that PCR technique is one of the useful tools in early and rapid diagnosis of pulmonary tuberculosis and silicotuberculosis.

Aged↗
PubMed1993

[The diagnosis of tuberculous meningitis using polymerase chain reaction].

The PCR system can detect 10 pg of DNA and 10-50 cfu of M. tuberculosis. The sensitivity and specificity rate of our PCR system about 80 cases of tuberculosis meningitis and 63 cases of nontuberculous meningitis is 85% and 98.4%, respectively. Comparing with the routine mycobacterial examination in 61 cases of tuberculous meningitis, the positive rate of PCR is 86.9%, while the positive rate of routine mycobacterial examination (smear and/or culture) is only 13.1%. In the 53 negative cases of routine mycobacterial examination, the positive rate of PCR can reach 86.8%. The whole detection can be finished within 2 days.

DNA, Bacterial↗
PubMed1992

[Studies on whole chromosomal DNA probes from Mycobacterium tuberculosis and Mycobacterium intracellulare].

The whole chromosomal DNA probes prepared from M. tuberculosis and M. intracellulare were used to hybridize in DNA dot blot and bacterial dot blot hybridization with 9 reference strains of Mycobacteria, including M. tuberculosis, M. bovis, M. avium, M. intracellulare, M kansasii, etc. These two probes showed good sensitivity and specificity for M. tuberculosis complex and M. avium complex respectively. Using the colony hybridization in situ, we can detect and identify M. tuberculosis with the probes on NC membrane after 1-2 week incubation over 7H9 agar plate.

Blotting, Southern↗
PubMed1991

Structure and expression of subunit A from the bovine chromaffin cell vacuolar ATPase.

Subunit A of the vacuolar H(+)-ATPase class is thought to be responsible for the ATP hydrolysis which drives proton-pumping. We report here the cloning and sequence determination of the first mammalian cDNA encoding a bovine vacuolar ATPase subunit A from an adrenal medulla cDNA library. Northern blots of bovine adrenal medulla RNA reveal a message of approximately 3.8 kb. The predicted peptide sequence, consisting of 618 amino acids with a calculated molecular weight of 68397 daltons, is similar to the sequences of the three known subunit A proteins. beta-Galactosidase-subunit A fusion proteins were immuno-decorated by an antiserum raised to the subunit A protein from corn coleoptile vacuoles.

Adenosine Triphosphatases↗