%0 Journal Article
%T Antiserum Preparation and Specific Detection of Banana RING-E3 Ubiquitin-Protein Ligase
%A Naitong Yu
%A Qin Zhou
%A Hongwei Yuan
%A Shuli Xian
%A Jianhua Wang
%A Yi Yang
%A Zhixin Liu
%J American Journal of Plant Sciences
%P 371-379
%@ 2158-2750
%D 2022
%I Scientific Research Publishing
%R 10.4236/ajps.2022.133022
%X According to the data of
banana transcriptome sequencing, an E3 ubiquitin-protein ligase gene was cloned
by RT-PCR method using the cDNA sample of banana leaves. The complete ORF of <i>E3
ubiquitin-protein ligase </i>is 681 bp long and its encoded protein showed 100%
sequence identity to homologue RING-H2 finger protein (XP_009407047.1) of
Musa_acuminata. Bioinformatic analysis indicated that E3 ubiquitin-protein
ligase contains the Ring finger domain in C terminus and eight cross-brace
motifs are found in the domain. The target gene was digested by EcoR V and EcoR
I, and was inserted into prokaryotic expression vector pET-32a of the same
digestions to obtain the plasmid pET32a-E3 ubiquitin-protein ligase. The
recombinant plasmid was introduced into <i>Escherichia coli</i> strain BL21
(DE3), and induced at 25<span style=\"font-family:;\" \"=\"\">ˇăC with 0.4 mmol/L
IPTG for 6 hours. The soluble fusion protein was expressed and high purity fusion
protein was obtained by Ni<sup>2+</sup>-NTA agarose affinity chromatography
purification. The fusion protein was injected into mice 3 times to prepare the
antiserum. Western blot analysis showed a specific protein band was detected in
total protein sample of banana leaves, but not for the samples of wild-type <i>Nicotiana
benthamiana</i> (N.B.) and wild-type <i>Arabidopsis thaliana</i> (A.T.),
implying the antiserum was specific to banana E3 ubiquitin-protein ligase.</span>
%K Banana
%K E3 Ubiquitin-Protein Ligase
%K Antiserum Preparation
%K Polyclonal Antibody
%U http://www.scirp.org/journal/PaperInformation.aspx?PaperID=115947