| Autor: |
Dimitrova, M., Denkova, L., Balinska, M., Peykov, S., Dimov, S. G. |
| Předmět: |
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| Zdroj: |
Genetics & Plant Physiology; 2018, Vol. 8 Issue 1/2, p51-61, 11p |
| Abstrakt: |
Two novel plasmid cloning vectors, pBlue and pYellow, were constructed by substituting the LacZ' gene fragment of pUC19 with amilCP and amilGFP chromoprotein coding sequences obtained from the coral Acropora millepora. Both sequences were codonoptimized for expression in Escherihia coli and, when expressed, produced blue and yellow coloring, respectively. The substitution of the LacZ' gene fragment eliminates the need for adding X-Gal to the growth medium while keeping the same mode for positive transformants selection as the one used by the ancestral vector. As a proof of the concept, a 644 bp long fragment from the dia-1 gene in D. melanogaster was cloned in our new vectors pBlue and pYellow using standard restriction and ligation techniques. Positive transformants were selected using the white color of the colonies as an indicator for a successful cloning event. All colonies that contained empty vectors developed the corresponding blue or yellow coloring in the process of cultivation, allowing their visual identification. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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