One-step Sequence- and Ligation-independent Cloning As a Rapid and Versatile Cloning Method for Functional Genomics Studies

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2012 May 22

PMID 22610439

Citations 201

Authors

Jae-Yeon Jeong

Hyung-Soon Yim

Ji-Young Ryu

Hyun Sook Lee

Jung-Hyun Lee

Dong-Seung Seen

Sung Gyun Kang

Affiliations

Soon will be listed here.

Abstract

We developed one-step sequence- and ligation-independent cloning (SLIC) as a simple, cost-effective, time-saving, and versatile cloning method. Highly efficient and directional cloning can be achieved by direct bacterial transformation 2.5 min after mixing any linearized vector, an insert(s) prepared by PCR, and T4 DNA polymerase in a tube at room temperature.

Citing Articles

Selective Degradation of Cucumber Mosaic Virus RNA3 by Nonsense-Mediated Decay Benefits Viral Early Infection.

Zhao D, Ahmed M, Tian M, Li M, Gu Z, Liao Q Mol Plant Pathol. 2025; 26(3):e70070.

PMID: 40059085 PMC: 11890980. DOI: 10.1111/mpp.70070.

Expression of transgenic biotin ligases in inducible neuronal murine cell lines by integration into the mHipp11 gene locus.

Feicht L, Dangel A, Jansen R PLoS One. 2025; 20(3):e0315806.

PMID: 40036200 PMC: 11878913. DOI: 10.1371/journal.pone.0315806.

Regulation of CXCR4 function by S1P through heteromerization.

Kim H, Jeong J, Huh W Cell Commun Signal. 2025; 23(1):111.

PMID: 40012038 PMC: 11863771. DOI: 10.1186/s12964-025-02099-x.

Mechanism of Centrosomal Protein 55 (CEP55) Loading Into Exosomes.

Dahlstroem C, Barezani J, Li J, Sopelniak K, Muhs S, Schneider C J Extracell Vesicles. 2025; 14(2):e70046.

PMID: 39976236 PMC: 11840697. DOI: 10.1002/jev2.70046.

DAPE cloning with modified primers for producing designated lengths of 3' single-stranded ends in PCR products.

Lee S, Kim H, Aqsa , Jeung K, Won M, Ro H PLoS One. 2025; 20(2):e0318015.

PMID: 39946422 PMC: 11825038. DOI: 10.1371/journal.pone.0318015.

References

Aslanidis C, de Jong P . Ligation-independent cloning of PCR products (LIC-PCR). Nucleic Acids Res. 1990; 18(20):6069-74. PMC: 332407. DOI: 10.1093/nar/18.20.6069. View

Matsumi R, Manabe K, Fukui T, Atomi H, Imanaka T . Disruption of a sugar transporter gene cluster in a hyperthermophilic archaeon using a host-marker system based on antibiotic resistance. J Bacteriol. 2007; 189(7):2683-91. PMC: 1855824. DOI: 10.1128/JB.01692-06. View

Kuijper J, Wiren K, Mathies L, Gray C, Hagen F . Functional cloning vectors for use in directional cDNA cloning using cohesive ends produced with T4 DNA polymerase. Gene. 1992; 112(2):147-55. DOI: 10.1016/0378-1119(92)90370-5. View

Hileman T, Santangelo T . Genetics Techniques for Thermococcus kodakarensis. Front Microbiol. 2012; 3:195. PMC: 3370424. DOI: 10.3389/fmicb.2012.00195. View

Li M, Elledge S . Harnessing homologous recombination in vitro to generate recombinant DNA via SLIC. Nat Methods. 2007; 4(3):251-6. DOI: 10.1038/nmeth1010. View

Li C, Evans R . Ligation independent cloning irrespective of restriction site compatibility. Nucleic Acids Res. 1997; 25(20):4165-6. PMC: 147003. DOI: 10.1093/nar/25.20.4165. View

Xu R, Li Q . Protocol: Streamline cloning of genes into binary vectors in Agrobacterium via the Gateway(R) TOPO vector system. Plant Methods. 2008; 4:4. PMC: 2257932. DOI: 10.1186/1746-4811-4-4. View

Kuzminov A . Recombinational repair of DNA damage in Escherichia coli and bacteriophage lambda. Microbiol Mol Biol Rev. 1999; 63(4):751-813, table of contents. PMC: 98976. DOI: 10.1128/MMBR.63.4.751-813.1999. View

Zhu B, Cai G, Hall E, Freeman G . In-fusion assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations. Biotechniques. 2007; 43(3):354-9. DOI: 10.2144/000112536. View

10.

Hamilton M, Nuara A, Gammon D, Buller R, Evans D . Duplex strand joining reactions catalyzed by vaccinia virus DNA polymerase. Nucleic Acids Res. 2006; 35(1):143-51. PMC: 1802553. DOI: 10.1093/nar/gkl1015. View

11.

de Jong R, Daniels M, Kaptein R, Folkers G . Enzyme free cloning for high throughput gene cloning and expression. J Struct Funct Genomics. 2007; 7(3-4):109-18. DOI: 10.1007/s10969-006-9014-z. View

12.

Cheo D, Titus S, Byrd D, Hartley J, Temple G, Brasch M . Concerted assembly and cloning of multiple DNA segments using in vitro site-specific recombination: functional analysis of multi-segment expression clones. Genome Res. 2004; 14(10B):2111-20. PMC: 528927. DOI: 10.1101/gr.2512204. View

13.

Walhout A, Temple G, Brasch M, HARTLEY J, Lorson M, Van den Heuvel S . GATEWAY recombinational cloning: application to the cloning of large numbers of open reading frames or ORFeomes. Methods Enzymol. 2000; 328:575-92. DOI: 10.1016/s0076-6879(00)28419-x. View

14.

Zhang Y, Buchholz F, Muyrers J, Stewart A . A new logic for DNA engineering using recombination in Escherichia coli. Nat Genet. 1998; 20(2):123-8. DOI: 10.1038/2417. View

15.

Lee H, Kang S, Bae S, Lim J, Cho Y, Kim Y . The complete genome sequence of Thermococcus onnurineus NA1 reveals a mixed heterotrophic and carboxydotrophic metabolism. J Bacteriol. 2008; 190(22):7491-9. PMC: 2576655. DOI: 10.1128/JB.00746-08. View