Abstract：
BACKGROUND:Prevalence of fibroproliferative diseases, including chronic kidney disease is rapidly increasing and has become a major public health problem worldwide. Fibroproliferative diseases are characterized by increased expression of α smooth muscle actin (α-SMA) that belongs to the family of the six conserved actin isoforms showing high degree homology. The aim of the present study was to develop real-time PCRs that clearly discriminate α-SMA and ß-actin from other actin isoforms. RESULTS:Real-time PCRs using self-designed mouse, human and rat specific α-SMA or ß-actin primer pairs resulted in the specific amplification of the artificial DNA templates corresponding to mouse, human or rat α-SMA or ß-actin, however ß-actin showed cross-reaction with the housekeeping γ-cyto-actin. We have shown that the use of improperly designed literary primer pairs significantly affects the results of PCRs measuring mRNA expression of α-SMA or ß-actin in the kidney of mice underwent UUO. CONCLUSION:We developed a set of carefully designed primer pairs and PCR conditions to selectively determine the expression of mouse, human or rat α-SMA and ß-actin isoforms. We demonstrated the importance of primer specificity in experiments where the results are normalized to the expression of ß-actin especially when fibrosis and thus increased expression of α-SMA is occur.
journal_name
BMC Mol Bioljournal_title
BMC molecular biologyauthors
Veres-Székely A,Pap D,Sziksz E,Jávorszky E,Rokonay R,Lippai R,Tory K,Fekete A,Tulassay T,Szabó AJ,Vannay Ádoi
10.1186/s12867-017-0089-9subject
Has Abstractpub_date
2017-04-27 00:00:00pages
12issue
1issn
1471-2199pii
10.1186/s12867-017-0089-9journal_volume
18pub_type
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