ABCMdb

ABCC7 p.Glu217Arg

None has been submitted yet.

Publications

PMID: 10220334 [PubMed] Chen M et al: "Topogenesis of cystic fibrosis transmembrane conductance regulator (CFTR): regulation by the amino terminal transmembrane sequences."

No. Sentence Comment

47 Primers carrying various mutations are 5'-AATCTGGAGGTTGTTAAAGGCGTC-3' (E217R/Q220K), 5'-CATCATTTCCCCTAGCCC-3' (R242E), 5'-CT- GATCTTCGTAATTCATCATCAT-3' (K246N/R248E), and 5'-TCCCAGCTTCCTGATCT-3' (R251E). Login to comment
48 To make the CFTR-N4R(-8) and CFTR-N4R(-5) mutants, two PCR reactions were performed as described above using CFTR-N4R(E217R/Q220K) or wild-type CFTR-N4R as templates and the mutant primer 5'-AGGGGAAATGATGATGGAG- TACGAAGATCAGGAAGCTGGGGATATCAG-3'. Login to comment
49 The resulting constructs were named CFTR-N4R(E217R/Q220K), CFTR-N4R(R242E), CFTR-N4R(K246N/R248E), CFTR-N4R(R251E), CFTR-N4R(-8), and CFTR-N4R(-5). Login to comment
50 To remove TM1 and TM2 from CFTR-N4R, CFTR-N4R- (E217R/Q220K), CFTR-N4R(-8), and CFTR-N4R(-5), PCR was performed using a primer 5'-GAGACCATGCA- GATGAGAATAG-3' containing Kozak translation initiation codon and a primer 5'-CACTTTTGCCAACCAG-3' in the glycosylation reporter sequence on templates CFTR-N4R, CFTR-N4R(E217R/Q220K), CFTR-N4R(-8), and CFTR-N4R(-5), respectively. Login to comment
53 The final DNA clones were named CF-TM3,4R, CF-TM3,4R(E217R/Q220K), CF-TM3,4R(-8), and CF-TM3,4R(-5), respectively. To engineer R242E and K246N/R248E mutations into CF-TM3,4R, an EcoRI-EcoNI fragment encoding TM3 and part of TM4 was released from CF-TM3,4R and used to replace the amino terminal-encoding sequence in CFTR-N4R(R242E) and CFTR-N4R(K246N/R248E). Login to comment
72 The mutant molecules used were CF-TM3,4R(E217R/ Q220K), CF-TM3,4R(R242E), and CF-TM3,4R(K246N/ R248E). Login to comment