Cloning And Analysis On Bioinformatics Of Aspergillus Ubiquitin-conjugating Enzyme Gene

Salt stress is one of the major environmental factor which inhibits plant growth,reduces crop yields. For a long time, it becomes the focus of attention on how toimprove the salt tolerance of plants, to increase crop yield under salt stress. Based onthe above facts, cultivating a salt plant on the production of human life is imminent.This paper based on experimental salt Aspergillus ubiquitin-conjugating enzyme genecloning, analysis of the cloned gene, in order to contribute to the genetic engineeringof transgenic plants.The test conditions and stuffs of Aspergillums physiological identifications inthis paper, which provided by Jilin University College of Plant Science ZhangShihong’s laboratory in this paper. The experimental stuffs—Aspergillums have avery strong salt resistance, suggesting the multi-class gene of Aspergillums that maybe regulation or directly give the function of its salt. The ubiquitin-proteasomesystem plays an important role in the growth and development of biological process,which not only to ensure the normal physiological function of the plant, but alsomaintain the state of the plant for basic survival in the face of adversity.Ubiquitin-conjugating enzyme terminates the explanation of the target proteins andthe target proteins regulation. Ubiquitin-conjugating enzymes in cell metabolism animportant role in determining factor.Ubiquitin-conjugating enzyme functional domain cDNA sequences by libraryscreening, sequencing and bioinformatics analysis, suggesting that the cloned genefor ubiquitin-conjugating enzyme gene from Aspergillums ESTs were found. Theeukaryotic expression vector, the expected use of Agro bacterium-mediatedtransformation of Arabidopsis to infection so as to achieve the purpose of genetictransformation, and genetically pure and genetically modified plants.By bioinformatic analysis of the ubiquitin-conjugating enzyme gene got from thespeculating clone, we get the following information:1.The Aspergillus UBE2molecular weight size16.6ku, its basic amino acids (K, R),acidic amino acids (D and E), hydrophobic amino acids (A, I, L, and F, W, V) andpolar amino acids (N, C, Q, S, T, Y) content is15,16,45and41, the isoelectric pointis PH6.4. 2. The section62~64nt is the first codon of the cDNA sequence, the62~505nt ofthe Aspergillus UBE2sequence has a complete ORF, which encodes a147AA proteinsequence.3. Let the amino acid sequence deduced from the cloning of cDNA as a probe, forBlastP analysis to GenBank non-redundant protein database, we find Zea mays,Saccharomyces cerevisiae, Danio rerio,Griffithsiajaponica, Arabidopsis thaliana andBos taurus， the identity of ubiquitin-conjugating enzyme gene sequence is84%,83%,85%,88%,80%and83%.4. Predictions results of E2has α-helices accounting for33.33%,β extends accountingfor19.05%,47.62%random coil.5. Aspergillus E2conserved domain analysis, dplay of Aspergillus E2No.4to136belong to the ubiquitin-conjugating enzyme family, the74to89is activity ofubiquitin conjugating enzyme, and has the following conserved sequence[FYWLSP-H-[PC]-[NHL]-[LIV]-x (3,4)-Gx-[LIVP]-C-[LIV]-x (1,2)-[LIVR].Comprehensive bioinformatics analysis of the results, it’s hypothesized that theubiquitin-conjugating enzyme gene overexpression may be the stress response byregulating the activity of the target protein, thus the regulation of genetically modifiedplants.