Cacao

	GO:0003887	DNA-directed DNA polymerase activity	PMID:22108584	ECO:0001258 IDA: Inferred from Direct Assay		Molecular Function	The gp43 protein from the bacteriophage BFK20 of the Brevibacterium flavum was suspected to exhibit polymerase activity. The gp43 was isolated and its polymerase activity was quantified by the amount of released free pyrophosphate through a colorimetric spectrophotometer analysis in vitro. The reaction mixture contained ssDNA and dNTP, and the addition of gp43 started the reaction with single-stranded DNA used as the template. Aliquots from the mixture were then added to thermostable inorganic pyrophosphatase.The phosphate that was released was quantified through a colorimetric spectrophotometer analysis, and results were shown in Figure 4. A negative control consisted of the reaction mixture without the thermostable inorganic pyrophosphatase. The negative control was subtracted from the experimental values to obtain the amount of phosphate released. From this procedure, researchers concluded that the gp43 protein was able to synthesize DNA.	complete
This annotation made on page: 9CAUD:Q3V5F2 By: Shreya1 (group Team Purple B 2019) on 2019-04-10 10:32:18 CDT.

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Team Purple B 2019	5

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2019-04-20 04:52:20 CDT

great work!

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2019-04-18 16:34:51 CDT

This annotation has been flagged because it has been edited since last assessment

Qualifier	GO ID	GO term name	Reference	ECO ID	ECO term name	with/from	Aspect	Extension	Notes	Status
	GO:0003887	DNA-directed DNA polymerase activity	PMID:22108584	ECO:0001258	IDA: Inferred from Direct Assay		F		The gp43 protein from the bacteriophage BFK20 of the Brevibacterium flavum was suspected to exhibit polymerase activity. To purify samples of gp43, a plasmid that produces gp43 was transformed into E. coli, these cells were centrifuged, the supernatant was loaded onto a cobalt-affinity chromatography column, and the gp43 was collected from the column. The polymerase activity of gp43 was quantified by the amount of released free pyrophosphate through a colorimetric spectrophotometer analysis in vitro. The reaction mixture contained ssDNA and dNTP, and the addition of gp43 started the reaction with single-stranded DNA used as the template. Aliquots from the mixture were then added to thermostable inorganic pyrophosphatase.The phosphate that was released was quantified through a colorimetric spectrophotometer analysis, and results were shown in Figure 4. A negative control consisted of the reaction mixture without the thermostable inorganic pyrophosphatase. The negative control was subtracted from the experimental values to obtain the amount of phosphate released. From this procedure, researchers concluded that the gp43 protein was able to synthesize DNA.	complete CACAO 13720

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2019-04-17 23:26:46 CDT

Please reword more technically: "The gp43 was isolated"

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