alesio antonini experience

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Member Since: 5/4/2006
Band Members: Alesio Antonini, Experience. S. cerevisiae strains containing RAS2val19, a RAS2 gene with a missense mutation analogous to one that activates the transforming potential of mammalian ras genes, have growth and biochemical properties strikingly similar to yeast strains carrying IAC or bcy1. Yeast strains carrying the IAC mutation have elevated levels of adenylate cyclase activity. bcy1 is a mutation that suppresses the lethality in adenylate cyclase deficient yeast. Yeast strains deficient in RAS function exhibit properties similar to adenylate cyclase deficient yeast. bcy1 suppresses lethality in ras1-ras2- yeast. Compared to wild-type yeast strains, intracellular cyclic AMP levels are significantly elevated in RAS2val19 strains, significantly depressed in ras2- strains, and virtually undetectable in ras1-ras2-bcy1 strains. Membranes from ras1-ras2-bcy1 yeast lack the GTP-stimulated adenylate cyclase activity present in membranes from wild-type cells, and membranes from RAS2val19 yeast strains have elevated levels of an apparently GTP-independent adenylate cyclase activity. Mixing membranes from ras1-ras2- yeast with membranes from adenylate cyclase deficient yeast reconstitutes a GTP-dependent adenylate cyclase.An important recent advance in the functional analysis of Saccharomyces cerevisiae genes is the development of the one-step PCR-mediated technique for deletion and modification of chromosomal genes. This method allows very rapid gene manipulations without requiring plasmid clones of the gene of interest. We describe here a new set of plasmids that serve as templates for the PCR synthesis of fragments that allow a variety of gene modifications. Using as selectable marker the S. cerevisiae TRP1 gene or modules containing the heterologous Schizosaccharomyces pombe his5+ or Escherichia coli kanr gene, these plasmids allow gene deletion, gene overexpression (using the regulatable GAL1 promoter), C- or N-terminal protein tagging [with GFP(S65T), GST, or the 3HA or 13Myc epitope], and partial N- or C-terminal deletions (with or without concomitant protein tagging). Because of the modular nature of the plasmids, they allow efficient and economical use of a small number of PCR primers for a wide variety of gene manipulations. Thus, these plasmids should further facilitate the rapid analysis of gene function in S. cerevisiae.
Influences: Monowatt,Dilatazione,sfingolipide,tetranicus urticae,Yatanawal Nuri Buzid fi hadhihi al-maqalah al-mutarjamah ila al-Injiliziyah [al-sadirah fi al-tariq, al-sinah 47, al-'adad 4 (Aylul Sibtimbir 1988)] mawdu' al-waqi'iyah al-jadidah fi al-sinima al-'Arabiyah. Wa-allati yad'u ilayha ka-badil lima huwa rahin. Wa-fima yali muqtatifat min al-maqalah allati turjimat bil-kamil ila al-injiliziyah fi hadha al-'adad: "Rubbama la takfi kalimat "hazimah" lil-ta'bir 'an wad' al-'alam al-'Arabi, li-anna al-azmah allati yatakhabbat fiha hiya a'maq min an takun murtabitah bi-hazimat 1967 li-annaha laysat azmah zarifah aw mutamarkizah fi balad dun al-akhar bi-qadr ma hiya azmah tarikhiyah, haykaliyah, mutajadhdhirah wa-tarja' usuluha ila al-inhitat al-hadari al-ladhi 'ashathu al-buldan al-'Arabiyah al-Islamiyah mundhu qurun 'adidah." "Tu'tabar aflam "al-Mumya'",saccharomyces cerevisiae "al-Mukhdu'un" wa "al-'Asfur" thalathah ibda'at mujaddidah tamaman fi madmuniha wa-shakliha... "al-Mumya'" yabda' tajribah ra'idah fi mawdu' asbaha simmah min simmat al-waqi'yah al-jadidah wa-huwa al-bahth 'an "al-hawiyah" wa-al-'ilaqah bil-turath wa bil-shakhsiyah... wa-ya'ti "al-Mukhdu'un" tribolium castaneum li-tawfiq Salih ka-tatwij li-a'mal al-mukhrij al-sabiqah allati hawala fiha al-qat' ma'a al-miludrama wa-lakin lam yakun al-qat' niha'iyan illa fi "al-Mukhdu'un". Fa-huwa yuqaddim khafaya al-qadiyah al-Filastiniyah wa al-wajh al-'Arabi li-taqtil al-Filastiniyyin... Amma film "al-'Usfur" fa-huwa qimmah al-sinima al-'Arabiyah 'ala al-itlaq. Jadid fi mawdu'uhi 'ala al-mustawa al-'alami... Hadhihi al-aflam al-thalathah tajtami' fi niqtah hammah: Inna al-insan al-basit makhdu' min taraf al-sultah, wa-hazimatahu tukmin fi annahu sallama al-sultah li-ghayrih, wa-qabl al-wasayah." "Amma al-jil al-jadid min al-sinima'iyin al-ladhina wasalu masirat Tawfiq Salah wa Shahin wa Shadi 'Abd al-Salam fahum fi aghlabihim, in lam yakunu kullahum, min kharij Misr. Wa-bil-tahdid min buldan al-Maghrib al-'Arabi wa Suriyya wa Lubnan cazo in culo wa Falistin wa-aghlabahum min mawalid al-arba'inat". "Wa-min khususiyat al-waqi'iyah al-jadidah al-bahth 'an al-hawiyah allati asbahat 'inda al-'adid niqtat istifham. Wa al-bahth 'an al-hawiyah ya'ni al-bahth 'an al-shakhsiyah wa al-bahth 'an al-madi al-majid al-muzdahir amilan fi irja'ihi... [wa-aydan] min khususiyat al-waqi'iyah al-jadidah tahtim al-akadhib wa al-usturah allati banaha al-Adab al-qadim wa al-sinima al-qadimah". "Wa-idha aradna talkhis al-waqi'iyah al-jadidah fi jumlah naqul: "Khalq al-jadid ma'a al-jamil". Laysat hunak namadhij wa-la kalishihat tataba', bal tatahaddad al-mawadi' wa al-turuq bi-ta'addud al-mukhrijin." "al-Sinima al-jadidah mushattatah, muba'tharah, aghlabaha dafin al-sanadiq wa al-'ulab, al-kullu yarid an yata'arraf 'alayha bidun an yashtariha, kullu sinah tazdad su'ubah tamwilaha, hatta al-i'anat allati kana yahsal 'alayha al-mukhrijun min Urubba taqallasat... al-hal fi aydina wa-laysa fi ayyi namudhaj wa-la itijah."
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Record Label: We have identified mutant strains of S. cerevisiae
Type of Label: Major