Difference between revisions of "Part:BBa K1639015"

Revision as of 16:44, 16 September 2015 (view source) Registry (Talk | contribs)		Latest revision as of 23:15, 21 September 2015 (view source) Nurish (Talk | contribs) (→‎Usage and Biology)
(5 intermediate revisions by the same user not shown)
Line 1:		Line 1:
−
	__NOTOC__		__NOTOC__
	<partinfo>BBa_K1639015 short</partinfo>		<partinfo>BBa_K1639015 short</partinfo>
−	a
−	<!-- Add more about the biology of this part here	+
		+
	===Usage and Biology===		===Usage and Biology===
		+	'''Cloning into pTet-off'''
		+	This part contains VP16 activator complex with appropriate[[File:pTet off-373.png|right|360px|thumb|'''Figure 1:'''Schematic diagram of cloning into pTet-off]] enzyme cut sites and miRNA binding sites for high-miRNas. Originally it's same with VP16 part of tetracycline-controlled transactivator (tTA), comprising a fusion of the '''tetracycline repressor TetR with the C-terminal activation domain of herpes simplex virus VP16''' By including Sal1 cut site at the beginning of this part we can change it with original tTA part found in pTet-off plasmid. ''(Figure 1)'' We performed ligation and then transformed the plasmids into BL21 bacteria. After 16 hours incubation at 37 C, we performed colony PCR and cut-check to check for positive clones. Despite trying this strategy 7 or 8 times, we were unable to successfully clone this g-block into pTET off. <br clea=all>
		+
		+	Below there is a whole diagram of Cancer module. This part is first step in this machinery. ''(Figure 2)''
		+	[[File:ATOMS-Turkiye_cancer_switch_1.1.png|center|450px|thumb|'''Figure 2''']]
		+
		+
	<!-- -->		<!-- -->

---

Latest revision as of 23:15, 21 September 2015

VP16 Transcriptional Activation Domain with miR373 Binding Site

Usage and Biology

Cloning into pTet-off

This part contains VP16 activator complex with appropriate enzyme cut sites and miRNA binding sites for high-miRNas. Originally it's same with VP16 part of tetracycline-controlled transactivator (tTA), comprising a fusion of the tetracycline repressor TetR with the C-terminal activation domain of herpes simplex virus VP16 By including Sal1 cut site at the beginning of this part we can change it with original tTA part found in pTet-off plasmid. (Figure 1) We performed ligation and then transformed the plasmids into BL21 bacteria. After 16 hours incubation at 37 C, we performed colony PCR and cut-check to check for positive clones. Despite trying this strategy 7 or 8 times, we were unable to successfully clone this g-block into pTET off.

Below there is a whole diagram of Cancer module. This part is first step in this machinery. (Figure 2)

Sequence and Features