Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (8): 135-141.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1509
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SUO Qing-qing(), WU Nan, YANG Hui, LI Li(), WANG Xi-feng
Received:
2021-12-06
Online:
2022-08-26
Published:
2022-09-14
Contact:
LI Li
E-mail:15093157487@163.com;lli@ippcaas.cn
SUO Qing-qing, WU Nan, YANG Hui, LI Li, WANG Xi-feng. Prokaryotic Expression,Antibody Preparation and Application of Rice Caffeoyl Coenzyme A-O-methyltransferase Gene[J]. Biotechnology Bulletin, 2022, 38(8): 135-141.
Fig.2 Restriction identification of recombinant plasmid pET30a(+)-OsCCoAOMT1 M:DL 5000 DNA marker;1:pET30a(+)-OsCCoAOMT1 digested by Nde I and Hind III;2:pET30a(+)plasmid
Fig.3 SDS-PAGE analysis of recombinant plasmid pET30a(+)-OsCCoAOMT1 under the different inducing condition M:Protein marker;PC1:BSA(1 μg);PC2:BSA(2 μg). NC:Cell lysate without induction. 1:Cell lysate with induction for 16 h at 15℃. 2:Cell lysate with induction for 4 h at 37℃. NC1:Supernatant of cell lysate without induction. 3:Supernatant of cell lysate with induction for 16 h at 15℃. 4:Supernatant of cell lysate with induction for 4 h at 37℃. NC2:Pellet of cell lysate without induction. 5:Pellet of cell lysate with induction for 16 h at 15℃. 6:Pellet of cell lysate with induction for 4 h at 37℃
Fig.4 Western blot analysis of recombinant plasmid pET30a(+)-OsCCoAOMT1 under the different inducing condition M:Western blot marker. 1:Supernatant of cell lysate induced at 15℃ for 16 h. 2:Supernatant of cell lysate induced at 37℃ for 4 h
Fig.6 Western blot analysis of OsCCoAOMT1 in rice stem and leaf M:Western blot marker. A:Rice control antibody(1:rice stem protein;2:rice leaf protein). B:OsCCoAOMT1 antibody(1:rice stem protein;2:rice leaf protein)
Fig.7 Western blot analysis of different protein concentra-tions in rice M:Western blot marker;A:rice control antibody(1:100 μg;2:50 μg;3:25 μg;4:12.5 μg). B:OsCCoAOMT1 antibody(1:100 μg;2:50 μg;3:25 μg;4:12.5 μg)
Fig. 9 Expression level analysis the OsCCoAOMT1 protein in rice cytoplasm and nucleus M:Western blot marker;1:rice stem cytoplasmic protein;2:rice stem nucleoprotein
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