Cloning and Expression of Actin Gene Fragment in Hordeum Brevisubulatum
Wild barley(Hordeum brevisubulatum)is an important gramineous forage.It has a variety of stress resis-tance.To further explore and study the stress resistance genes of H.brevisubulatum,this study cloned the core fragment of the H.brevisubulatum actin gene(ACT)using RT-PCR method.Its sequence and expression characteristics were ana-lyzed by BLAST sequence alignment and qRT-PCR real-time quantitative method.The results showed that the ACT gene fragment of H.brevisubulatum was 518 bp in length and encoded 171 amino acids.ACT gene fragment of H.brevi-subulatum was 97%consistent with the nucleotide sequence of Hordeum vulgare ACT gene,and the amino acid se-quence homology of ACT protein was 91%by through BLAST analysis.The actin gene fragment of H.brevisubulatum was identified and named HbACT.The amino acid sequences of HbACT were compared with those of other 11 plants,and 152 conserved amino acids and 19 non-conserved amino acids were found,which proved that HbACT protein was highly conserved.qRT-PCR analysis showed that the mean Ct value of HbACT in the organs of H.brevisubulatum treated with different salt concentrations and salt treatment time was 19.30,the coefficient of variation in the ground part was 3.5,and the coefficient of kurtosis was 0.262.The variation coefficient and kurtosis coefficient of the under-ground part were 5.3 and 0.409,both belonging to the peak distribution.The results of RefFinder comprehensive analy-sis showed that the stability coefficient of Ct value was less than 1.7.In conclusion,HbACT gene expression was stable.It could be used as an internal reference gene to study the expression pattern analysis of functional genes in H.brevisu-bulatum.
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