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The double insertion events in Tragelaphini obviously took place after Bov-A2 inserted to Antilopinae' TP53P1 promoter, around 14MYA, as Bovini members (the cow/buffalo tribe) lack Bov-A2 in TP53 promoterter [ 43].
Firstly, double insertion lines accumulated free Asn.
The characteristics of these double insertion lines could be summarized in two points.
Recently, Arabidopsis T-DNA insertion lines for ASNase1 and ASNase2 were isolated, respectively, and then, two insertion lines were crossed, and double insertion lines were generated (Ivanov et al. 2012).
The double insertion homozygous diplotype was observed in 16.33% of the JBr animals and in 5.00% of the JB animals.
Sequencing revealed a double insertion of the oligo resulting in a triple strep-tag, which showed higher affinity compared to single tag.
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They also created reporter and conditional KI alleles with site-specific insertions of a short Tag or a long fluorescent reporter, and double insertions of two loxP sites (floxed alleles), respectively27.
Single insertions of 5 bp at nucleotides 7998/7999, or double insertions of 5 bp at nucleotides 7998/7999 and nucleotides 8197/8198, have been reported in flanking sequences of chloroplast encoding ORF, ORF100 (Takahashi et al. 2008).
In the partial B. anynana gDNA clone BA-ATG1 (GenBank: AY785062), double insertions by BaRTE elements were detected at both 5' and 3' termini.
Navarre, D., Parlier, A., Rudler, H. & Daran, J. C. Double acetylene insertion into a μ-methylene complex of iron: formation of a μ- divinyl carbene complex of iron after double hydrogen migration.
The transformed products were obtained as a result of de novo stereoselective reduction of the enone system, isomerization of double bond, insertion of double bond and hydroxylation.
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