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In a 1945 study by Demerec and Fano, [4] T7 was used to describe one of the seven phage types (T1 to T7) that grow lytically on Escherichia coli. [5] Although all seven phages were numbered arbitrarily, phages with odd numbers, or T-odd phages, were later discovered to share morphological and biochemical features that distinguish them from T-even phages. [6]
In biotechnology applications, T7 RNA polymerase is commonly used to transcribe DNA that has been cloned into vectors that have two (different) phage promoters (e.g., T7 and T3, or T7 and SP6) in opposite orientation. RNA can be selectively synthesized from either strand of the insert DNA with the different polymerases.
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Once this has occurred, the prohead undergoes maturation by cleavage of capsid subunits to form an icosahedral phage head with 5-fold symmetry. After the head maturation, the tail is joined in one of two ways: Either the tail is constructed separately, and joined with the connector, or the tail is constructed directly onto the phage head.
T7 DNA polymerase catalyzes the phosphoryl transfer [7] during DNA replication of the T7 phage. As shown in Figure 2, the 3’ hydroxyl group of a primer acts as a nucleophile and attacks the phosphodiester bond of nucleoside 5’-triphosphate (dTMP-PP). This reaction adds a nucleoside monophosphate into DNA and releases a pyrophosphate (PPi).
T7 DNA helicase (gp4) is a hexameric motor protein encoded by T7 phages that uses energy from dTTP hydrolysis to process unidirectionally along single stranded DNA, separating the two strands as it progresses. It is also a primase, making short stretches of RNA that initiates DNA synthesis. [2] It forms a complex with T7 DNA polymerase.
Podoviridae was a family of bacteriophage in the order Caudovirales often associated with T-7 like phages. [1] The family and order Caudovirales have now been abolished, with the term podovirus now used to refer to the morphology of viruses in this former family. [2]
Soon, the lab was able to clone the T7 RNA polymerase and use it, along with the powerful T7 promoter, to transcribe copious amounts of almost any gene. [4] The development of the T7 expression system has been considered the most successful biotechnology developed at the Brookhaven National Laboratory, being licensed by over 900 companies which ...