In-Frame Cloning, Research Paper Example

In-frame cloning Using Restriction Enzyme

In-frame cloning allows the scientist researchers in total control of the nucleotide sequence, codon usage, promoter, and other regulatory elements, additionally, it assists in the placement of the unique restriction sites which usually occur throughout the structure of the recombinant (Marillonnet et al., 2020). The in-frame cloning method allows the flexible and precise construction of the synthetic genes by applying the standard recombinant techniques. The method also tends to be less expensive and materials are always readily available. The mRNA biological translation of producing peptide chain containing amino acids is arrived through reading the sequence of the nucleotide in terms of groups which may include triplets or codons (Katrolia et al., 2020). The codon reading can have a significant impact on the subsequent amino acid sequence. Reading below is known as frameshift because the reading is moved along the sequence. The difference in the peptide sequence, therefore the folding and protein properties tend to be altered in the process. Though in some scenario the results of the frameshift usually produce an immature stop codon leading to truncation of the protein. The regulation of the frame start from the starting point of the codon ‘’AUG’ this process also eases coding of the amino acid. The figure below shows the possible option of the cloning:

AseI – ATTTAAT

TAATTA

Ndel – CATATG

GTAGTC

SnaBI – TACGTA

ATGCAT

References

Marillonnet, S., & Grützner, R. (2020). Synthetic DNA Assembly Using Golden Gate Cloning and the Hierarchical Modular Cloning Pipeline. Current protocols in molecular biology, 130(1), e115.

Katrolia, P., Liu, X., Zhao, Y., Kopparapu, N. K., & Zheng, X. (2020). Gene cloning, expression, and homology modeling of first fibrinolytic enzyme from mushroom (Cordyceps militaris). International journal of biological macromolecules, 146, 897-906.