http://hdl.handle.net/10027/9 UIC College of Liberal Arts and Sciences Sun, 19 May 2013 08:36:51 GMT 2013-05-19T08:36:51Z http://hdl.handle.net/10027/9820 Islamic Feminism in India: Indian Muslim Women Activists and the Reform of Muslim Personal Law Vatuk, Sylvia I describe here a nascent ‘Islamic feminist’ movement in India, dedicated to the goal of achieving gender equity under Muslim Personal Law. In justifying their demands, these women activists refer neither to the Indian Constitution nor to the universalistic human rights principles that guide secular feminists campaigning for passage of a gender-neutral uniform civil code of personal law, but rather to the authority of the Qur’an—which, they claim, grants Muslim women numerous rights that in practice are routinely denied them. They accuse the male ‘ulama of foisting ‘patriarchal’ interpretations of the Qur’an on the unlettered Muslim masses and assert their right to read the Qur’an for themselves and interpret it in a woman-friendly way. Their activities reflect an increasing ‘fragmentation of religious authority’ in the globalizing Muslim world, associated with the spread of mass education, new forms of media and transport and a mobile labour force, in which clerical claims to exclusive authoritative knowledge are being questioned by a wide variety of new voices, women’s among them. Whether it can ultimately succeed is an open question but the movement is clearly having an impact, even on the clerical establishment itself, insofar as the legal issues it considers most pressing for women are concerned. Modern Asian Studies 42,2/3(2008) pp. 489–518. © 2007 Cambridge University Press doi:10.1017/S0026749X07003228 Tue, 01 Jan 2008 06:00:00 GMT http://hdl.handle.net/10027/9820 2008-01-01T06:00:00Z http://hdl.handle.net/10027/9810 The Effect of School Rape-Supportive Norms on Rape Proclivity Rape prevention programs have recently begun using social norms interventions in addition to, or in lieu of, individual-level interventions. These programs assume that rape-supportive social norms influence the likelihood of rape. The current study tests that assumption by analyzing how school-level aggregates of men‘s rape myth acceptance (RMA) and hostile masculinity affect rape proclivity. Data for this study come from 1326 male students in 11 high schools throughout Illinois. At the individual level, risk and protective factors were similar to past studies: higher RMA and hostile masculinity were associated with increases in rape proclivity. Conversely, believing men have a responsibility to prevent rape, that they would personally intervene to prevent assault, and that there are negative consequences for perpetrating rape were all associated with decreased rape proclivity. After controlling for these individual factors, results indicate that higher school social norms for hostile masculinity increase the odds of reporting some likelihood of sexual assault. Against hypotheses, school social norms for RMA did not have a direct negative effect on proclivity; however, these results were partially qualified by interactions. School social norms for RMA appear to affect students differently depending on their own RMA. Results support efforts to target both individual and community-level factors. Implications for prevention programs are discussed. Thu, 21 Feb 2013 06:00:00 GMT http://hdl.handle.net/10027/9810 2013-02-21T06:00:00Z http://hdl.handle.net/10027/9802 Generation of FN3 Monobodies That Selectively Bind to the Src Family of Protein Kinases Protein kinases, known for phosphorylating their protein substrates to relay signaling events in cells, contain 518 members and encompass ~2% of human genes. One subgroup of the kinase family, Src family kinases (SFKs), including Blk, Fgr, Fyn, Hck, Lck, Lyn, Src, and Yes, have been studied for their roles in cell proliferation, migration, differentiation and survival. Because SFKs members have a similar overall structure with highly conserved sequences, currently available antibodies fail to distinguish the active form of one kinase from other SFKs. As SFKs often phosphorylate substrates with similar amino acid sequence, current biosensors fail to discriminate activation of one kinase from other SFKs. To generate affinity reagents that can be used to study the activation of individual member of SFKs, I used phage display technology for directed evolution of FN3 monobodies. To increase the efficiency of constructing a phage library of FN3 monobodies with Kunkel mutagenesis, I made three modifications to the previously published protocol. First, I incubated bacterial cells at 25°C instead of 37°C to achieve a 2- to 7-fold increase in the yield of the single-stranded DNA template. Second, with the introduction of restriction endonuclease sites into the diversified loops of the FN3 coding region, phage libraries with diversity up to 1010 and 99-100% recombinant were constructed. Finally, I designed a digestion- and ligation-free method for constructing secondary libraries, using DNA fragments amplified by error-prone and asymetric PCR as primer for conducting Kunkel mutagenesis. To demonstrate the efficiency of this improved method, I constructed two secondary libraries based on a FN3 monobody that bound to the active form of Pak1 kinase. Screening one of the mutagenic libraries isolated three variants that bound 2- to 4-fold tighter than the original clone. For generating affinity reagents to one member of the SFKs, Fyn tyrosine kinase, a phage-display library was screened for monobodies binding to the Fyn SH3 domain. The affinity selection identified three monobodies that bound selectively to the Fyn SH3 domain. One of the isolates, G9, bound exclusively to the Fyn SH3 domain out of 150 SH3 domains that I tested and had a dissociation constant (KD) of 166 ± 6 nM. Interestingly, while the G9 monobody lacks proline in its randomized loops, it bound at the same site on the SH3 domain as proline-rich ligands. The G9 monobody could be used to pull-down active recombinant Fyn kinase in vitro, demonstrating its potential as a highly selective probe for detecting active cellular Fyn kinase. To generate affinity reagents for another SFKs member, Lyn tyrosine kinase, another phage library was screened. Two isolates, TA1 and TA8, selectively bound to the Lyn SH3 domain out of 150 SH3 domains. Both TA1 and TA8, in the absence of a canonical PxxP motif in their binding loops, competed with a PxxP proline-rich peptide, Tip, for binding to Lyn SH3 domain. Due to the weak affinity of TA8 for Lyn SH3 (KD= ~5 µM), I used affinity maturation techniques to identify variants that bound > 8-fold tighter than the original TA8 clone. As a proof-of-concept experiment, one monobody was converted into a sensor that increased fluorescence upon binding to the Lyn SH3 domain in solution. Thu, 21 Feb 2013 06:00:00 GMT http://hdl.handle.net/10027/9802 2013-02-21T06:00:00Z http://hdl.handle.net/10027/9800 Directed Evolution of the Forkhead-associated Domain to Generate Anti-Phosphospecific Reagents While affinity reagents are valuable tools for monitoring protein phosphorylation and studying signaling events in cells, generating them through immunization of animals with phosphopeptides is expensive, laborious and time consuming. An attractive alternative is to use protein evolution techniques and isolate new anti-phosphopeptide binding specificities from a library of variants of a phosphopeptide-binding domain. To explore this strategy, we attempted to display on the surface of bacteriophage M13, the N-terminal Forkhead-associated domain (FHA1) of yeast Rad53p, which is a naturally occurring phosphothreonine (pT)-binding domain, and found it to be non-functional due to misfolding in the bacterial periplasm. To overcome this limitation, a library of FHA1 variants was constructed by mutagenic PCR and functional variants were isolated after three rounds of affinity selection with its pT peptide ligand. A hydrophobic residue at position 34 in the β1-strand was discovered to be essential for phage-display of a functional FHA1 domain. Additionally, by heating the phage library to 50ºC prior to affinity selection with its cognate pT peptide, we identified a variant (G2) that was ~8ºC more thermal stable than the wild-type domain. Using G2 as a scaffold, we constructed phage-displayed libraries of FHA1 variants and affinity selected for variants that bound selectively to seven pT peptides. These reagents are renewable and have high protein yields (~20-25 mg/L), when expressed in Escherichia coli. Thus, we have changed the specificity of the FHA1 domain and demonstrated that engineering phosphopeptide-binding domains is an attractive avenue for generating new anti-phosphopeptide binding specificities in vitro by phage-display. Thu, 21 Feb 2013 06:00:00 GMT http://hdl.handle.net/10027/9800 2013-02-21T06:00:00Z