Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtaine...Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtained by reverse transcription polymerase chain reaction (RT-PCR) and cloned into pMD18-T vector. The positive clone identified by PCR was sequenced. The sequencing result showed that the Actin gene fragment from lris lacteal var.chinensis Fisch.Koidz contained about 598 bp, encoding 199 amino acids. Homology comparison with Actin gene sequences of other plants in the GenBank showed that it shared over 82% nueleotide sequence homology and 90% amino acid sequence homology. It indicated that this was the Actin gene. Because of the stability expression ofActin gene, it usually cited as the internal reference to study the expression and regulation of foundation in other genes of lris lacteal var.chinensis Fisch.Koidz well.展开更多
基金Supported by the Postdoctoral Scientific Research Foundation of Heilongjiang Province(LBH-Q10144)the Natural Science Foundation of Heilongjiang Province(C201112)Northeast Agricultural University Doctoral Research Fund(200830)
文摘Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtained by reverse transcription polymerase chain reaction (RT-PCR) and cloned into pMD18-T vector. The positive clone identified by PCR was sequenced. The sequencing result showed that the Actin gene fragment from lris lacteal var.chinensis Fisch.Koidz contained about 598 bp, encoding 199 amino acids. Homology comparison with Actin gene sequences of other plants in the GenBank showed that it shared over 82% nueleotide sequence homology and 90% amino acid sequence homology. It indicated that this was the Actin gene. Because of the stability expression ofActin gene, it usually cited as the internal reference to study the expression and regulation of foundation in other genes of lris lacteal var.chinensis Fisch.Koidz well.