Molecular Biology Methods to Construct Recombinant Fibrous Protein.
Agarose gel electrophoresis
Polymerase chain reaction
Seamless cloning
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
2
9
2021
pubmed:
3
9
2021
medline:
7
1
2022
Statut:
ppublish
Résumé
Recombinant technologies are often used to synthesize fibrous proteins that are difficult to separate and extract in nature, such as spider silks and elastin. Although the recombination techniques can be diverse, PCR, gel electrophoresis, and seamless cloning, as the basic methods of molecular biology, have been widely used for constructing fibrous proteins' homologous recombinant plasmids. Considering that some readers of this book may not have a molecular biology background, in this chapter, we will introduce these three most used and effective recombination techniques. For PCR, we primarily introduce colony PCR, high-fidelity PCR, and overlap PCR, which are three kinds of the most used methods. In terms of seamless cloning, the detailed protocols of Gibson Assembly and Golden Gate Assembly are introduced. The introduction of this chapter is expected to provide a comprehensive methodological reference for the following chapters to introduce the recombination of specific fibroin proteins.
Identifiants
pubmed: 34472061
doi: 10.1007/978-1-0716-1574-4_12
doi:
Substances chimiques
Scleroproteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
123-135Informations de copyright
© 2021. Springer Science+Business Media, LLC, part of Springer Nature.
Références
Lai Y-T, King NP, Yeates TO (2012) Principles for designing ordered protein assemblies. Trends Cell Biol 22:653–661
doi: 10.1016/j.tcb.2012.08.004
Bozic S, Doles T, Gradisar H, Jerala R (2013) New designed protein assemblies. Curr Opin Chem Biol 17:940–945
doi: 10.1016/j.cbpa.2013.10.014
Patterson DP (2011) Symmetry assembled supramolecular protein cages: investigating a strategy for constructing new biomaterials. Proquest, Umi Dissertation Publishing, Ann Arbor, Michigan
King NP, Lai YT (2013) Practical approaches to designing novel protein assemblies. Curr Opin Struct Biol 23:632–638
doi: 10.1016/j.sbi.2013.06.002
Whyburn GP, Li Y, Huang Y (2008) Protein and protein assembly based material structures. J Mater Chem 18:3755–3762
doi: 10.1039/b807421f
Beying N, Schmidt C, Pacher M, Houben A, Puchta H (2020) CRISPR–Cas9-mediated induction of heritable chromosomal translocations in Arabidopsis. Nat Plants 6:638–645
doi: 10.1038/s41477-020-0663-x
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
doi: 10.1126/science.239.4839.487
Kieleczawa J (2006) DNA sequencing II: optimizing preparation and cleanup (Vol. 2). Jones and Bartlett Publishers, Burlington, Massachusetts
Higuchi R, Krummel B, Saiki RK (1988) A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res 16:7351–7367
doi: 10.1093/nar/16.15.7351
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, vol [prepared for Use in the CSH Courses on the Molecular Cloning of Eukaryotic Genes]. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Gibson DG, Young L, Chuang R-Y, Venter JC, Hutchison CA, Smith HO (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 6:343–345
doi: 10.1038/nmeth.1318
Engler C, Kandzia R, Marillonnet S (2008) A one pot, one step, precision cloning method with high throughput capability. PLoS One 3:e3647
doi: 10.1371/journal.pone.0003647
Golden Gate Assembly | NEB. Biolabs, New England. https://www.neb.com/applications/cloning-and-synthetic-biology/dna-assembly-and-cloning/golden-gate-assembly