Avtor/Urednik		Benčina, Mojca; Legiša, Matic
Naslov		Non-radioactive multiple reverse transcription-PCR method used for low abundance mRNA quantification
Tip		članek
Vir		Biotechnol Tech
Vol. in št.		Letnik 13
Leto izdaje		1999
Obseg		str. 865-9
Jezik		eng
Abstrakt		A non-radioactive multiple reverse transcription-PCR method for the relative quantitative analysis of low abundance mRNA is described. Chemiluminescent detection of cDNA products amplified by PCR and incorporating biotin-labelled dCTP was used for analysis of specific gene expression. After determining the optimal concentrations of RNA and cDNA, the tube-to-tube variations were minimised by transcription and amplification of the endogenous standard mRNA coding for protease C from Aspergillus niger, together with the transcript of interest. All the obtained data were relative values comparing to the internal standard pepC. By the method the difference in mRNA level between the two transformants with different pkaC gene copy number coding for cAMP-dependent protein kinase (PKA) catalytic subunit from A. niger was detected which was in accordance with the diferences in specific PKA enzymatic activity between both strains.
Deskriptorji		ASPERGILLUS NIGER RNA, MESSENGER CYCLIC AMP-DEPENDENT PROTEIN KINASES GENE EXPRESSION DNA, COMPLEMENTARY POLYMERASE CHAIN REACTION