Reverse-polarity PAGE for examining DNA binding domain phosphorylation.
نویسندگان
چکیده
1.Barik, S. and M.S. Galinski. 1991. “Megaprimer” method of PCR: increased template concentration improves yield. BioTechniques 10:489-490. 2.Colosimo, A., Z. Xu, G. Novelli, B. Dallapiccola, and D.C. Gruenert. 1999. Simple version of “megaprimer” PCR for site-directed mutagenesis. BioTechniques 26:870-873. 3.Datta, A.K. 1995. Efficient amplification using “megaprimer” by asymmetric polymerase chain reaction. Nucleic Acids Res. 23:45304531. 4.Giebel, L.B. and R.A. Spritz. 1990. Site-directed mutagenesis using a double-stranded DNA fragment as a PCR primer. Nucleic Acids Res. 25:4947. 5.Ling, M.M. and B.H. Robinson. 1997. Approaches to DNA mutagenesis: an overview. Anal. Biochem. 254:157-178. 6.Sarkar, G. and S.S. Sommer. 1990. The “megaprimer” method of site-directed mutagenesis. BioTechniques 8:404-407. 7.Smith, A.M. and K.P. Klugman. “Megaprimer” method of PCR-based mutagenesis: the concentration of megaprimer is a critical factor. BioTechniques 22:438-442.
منابع مشابه
Phosphorylation of BATF regulates DNA binding: a novel mechanism for AP-1 (activator protein-1) regulation.
BATF is a member of the AP-1 (activator protein-1) family of bZIP (basic leucine zipper) transcription factors that form transcriptionally inhibitory, DNA binding heterodimers with Jun proteins. In the present study, we demonstrate that BATF is phosphorylated in vivo on multiple serine and threonine residues and at least one tyrosine residue. Reverse-polarity PAGE revealed that serine-43 and th...
متن کاملDNA-binding polarity of human replication protein A positions nucleases in nucleotide excision repair.
The human single-stranded DNA-binding replication A protein (RPA) is involved in various DNA-processing events. By comparing the affinity of hRPA for artificial DNA hairpin structures with 3'- or 5'-protruding single-stranded arms, we found that hRPA binds ssDNA with a defined polarity; a strong ssDNA interaction domain of hRPA is positioned at the 5' side of its binding region, a weak ssDNA-bi...
متن کاملهمسانهسازی و بیان ناحیه اتصالی نوروتوکسین نوع B باکتری کلستریدیوم بوتولینوم در باکتری E. coli
Backgrounds & Aims: Botulism is caused by botulinum toxin. The best way to avoid the neurotoxin syndrome caused by BONT/B is to use recombinant vaccine made of BONT/B binding domain because binding domain has sufficient epitops to stimulate immune response. Materials & Methods: Initially BONT/B binding domain gene sequence were obtained from GenBank. Then the primers were designed. PCR react...
متن کاملp32 is a novel mammalian Lgl binding protein that enhances the activity of protein kinase Cζ and regulates cell polarity
Lgl (lethal giant larvae) plays an important role in cell polarity. Atypical protein kinase C (aPKC) binds to and phosphorylates Lgl, and the phosphorylation negatively regulates Lgl activity. In this study, we identify p32 as a novel Lgl binding protein that directly binds to a domain on mammalian Lgl2 (mLgl2), which contains the aPKC phosphorylation site. p32 also binds to PKCzeta, and the th...
متن کاملA mutational analysis of dishevelled in Drosophila defines novel domains in the dishevelled protein as well as novel suppressing alleles of axin.
Drosophila dishevelled (dsh) functions in two pathways: it is necessary to transduce Wingless (Wg) signaling and it is required in planar cell polarity. To learn more about how Dsh can discriminate between these functions, we performed genetic screens to isolate additional dsh alleles and we examined the potential role of protein phosphorylation by site-directed mutagenesis. We identified two a...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- BioTechniques
دوره 34 1 شماره
صفحات -
تاریخ انتشار 2003