Cloning, Expression Analysis And Construction OF Eukaryotic Expression Vector OF The Acyl-coenzyme A Binding Protein Gene In Maize

Maize is one of the most important crops in the world.Salt,drought,low temperature,high temperature,heavy metal toxicity and other abiotic stress become important factors affecting corn production,maize resistant gene screening and study on its function,not only can provide theoretical basis for transgenic breeding of maize,but also wide application space.The acyl-coenzyme A binding protein(acyl-CoA-binding protein,ACBP) in the plant acyl-coenzyme A especially long-chain acyl coenzyme A(long-chainfatty acyl-CoA esters,LCACoA) has a strong affinity,and thus play an important role in the process of long-chain acyl coenzyme A transporter.LCACoA is the intermediate product of fat metabolism in addition to being an important component of energy substances,or cell membrane.Study found that in the Arabidopsis ACBP6gene overexpression can significantly improve the cold tolerance in plants.Previous studies focused mainly on animal of acyl-coenzyme A binding protein functions,in the domestic plants of acyl-coenzyme A binding protein research is less,and the stress gene expression and gene expression in various tissues of study is less,corn ACBP expression and function study of reported.In this paper,the expression of the maize in the ZmACBP features and functionality analysis.The main results are as follows:1. Cloning and sequence analysis of ZmACBP gene:The use of homologous cloning methods,in the maize cDNA library clones obtained two segment sequences,respectively276bp and270bp,and using a bioinformatics approach to the translation of the protein basic physicochemical properties were predicted.The sequence of translation protein multiple sequence alignment revealed the experimentally obtained gene belongs to the ACBP gene family.2. ZmACBPl gene expression in Maize in content analysis:By fluorescence quantitative PCR method,the discovery of the ZmACBPl gene in Maize Seedlings Induced by low temperatures when the expression,along with the low temperature stress prolonged ZmACBPlgene expression was upregulated trend,under low temperature stress within24hours of ZmACBPlgene expression has been showed rising trend.In the seedlings of corn in different tissues of ZmACBPl gene expression analysis revealed expression amount higher than the root,leaf and stem.Not found in ZmACBPl,under high temperature,NaCl,dehydration conditions for the induction of. 3. Transformation of ZmACBPl gene in yeast:Construction of the success of this experiment a group of eukaryotic expression vectors,transformed yeast after PCR detection and restriction enzyme identification,screening out3groups of positive transformant,and saved the positive transformants containing yeast liquid,for further validation of ZmACBP1gene function provide support.