Construction of eukaryotic expression plasmid of human PRX3 and its expression in HEK-293FT cells / 华中科技大学学报（医学）（英德文版）

To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.

Distinguishing clonal apple rootstocks by isozymes banding patterns.

Molecular characterisation of clonal apple rootstocks using isozymes was carried out to identify isozyme polymorphism in seven clonal apple rootstocks and to identify the most characteristic and stable enzyme markers for each individual rootstock. Five enzyme systems were studied out of which polyphenol oxidase, malate dehydrogenase, acid phosphatase and peroxidase were useful in discriminating among the rootstocks. The peroxidase enzyme system showed maximum variation and esterase showed the least variation among the rootstocks. Out of seven rootstocks, three were distinguished on the basis of one enzyme system only (M.3 with MDH or PER, M.7 with PPO or PER and MM. 111 with MDH). Out of the sixteen loci studied seven were found to be polymorphic. Genetic variation among the rootstocks was explained on the basis of various parameters. The percentage of polymorphic loci varied from 13.33 to 35.71 per cent.

Evaluation of katG codon 315 mutations among isoniazid sensitive and resistant mycobacterium tuberculosis isolates from Thailand.

Nine isoniazid (INH)-susceptible and 11 INH-resistant Mycobacterium tuberculosis clinical isolates were analyzed for katG codon 315 mutations by polymerase chain reaction (PCR) assay using primers MYC-32 and MYC-33, followed by restriction fragment length polymorphism. After AciI digestion of PCR products, all 9 INH-susceptible isolates and 5 out of 11 (45%) INH-resistant isolates showed 0.12 kb band, which was previously reported to indicate wild type, whereas, 6 of 11 (55%) INH-resistant isolates lacked this band.

Kat G mutations in isoniazid resistant Mycobacterium tuberculosis isolates from Thai patients.

Isoniazid resistant mechanisms in Mycobacterium tuberculosis have been shown to involve at least two genes, kat G and inh A. Alteration in the kat G gene has been found in a great number of resistant isolates. Percentage of resistant isolates harboring alteration in this gene varied among laboratories suggesting that different mutations were presented in different geographic areas. Fourteen isoniazid resistant and five multidrug resistant isolates from the Central Chest Hospital, Thailand, were examined for the kat G gene mutations in the region between base position 17 to 299. No different pattern of mutations were found between these two groups. Among nineteen isolates, there were nine isolates which showed point mutations and five isolates with base insertions of the kat G gene. The remaining five isolates revealed gene deletion. Heteroduplex formation technique also confirmed base alterations in these nine mutants.