Biochemical Characterization of Nonamer Binding Domain of RAG1 Reveals its Thymine Preference with Respect to Length and Position

RAG complex consisting of RAG1 and RAG2 is a site-specific endonuclease responsible for the generation of antigen receptor diversity. It cleaves recombination signal sequence (RSS), comprising of conserved heptamer and nonamer. Nonamer binding domain (NBD) of RAG1 plays a central role in the recognition of RSS. To investigate the DNA binding properties of the domain, NBD of murine RAG1 was cloned, expressed and purified. Electrophoretic mobility shift assays showed that NBD binds with high affinity to nonamer in the context of 12/23 RSS or heteroduplex DNA. NBD binding was specific to thymines when single stranded DNA containing poly A, C, G or T were used. Biolayer interferometry studies showed that poly T binding to NBD was robust and comparable to that of 12RSS. More than 23 nt was essential for NBD binding at homothymidine stretches. On a double-stranded DNA, NBD could bind to A:T stretches, but not G:C or random sequences. Although NBD is indispensable for sequence specific activity of RAGs, external supplementation of purified nonamer binding domain to NBD deleted cRAG1/cRAG2 did not restore its activity, suggesting that the overall domain architecture of RAG1 is important. Therefore, we define the sequence requirements of NBD binding to DNA.

Analysis of sine-triangle and zero-sequence injection modulation schemes for split-phase induction motor drive

A split-phase induction motor is fed from two three-phase voltage source inverters for speed control. This study analyses carrier-comparison based pulse width modulation (PWM) schemes for a split-phase motor drive, from a space-vector perspective. Sine-triangle PWM, one zero-sequence injection PWM where the same zero-sequence signal is used for both the inverters, and another zero-sequence injection PWM where different zero-sequence signals are employed for the two inverters are considered. The set of voltage vectors applied, the sequence in which the voltage vectors are applied, and the resulting current ripple vector are analysed for all the PWM methods. Besides all the PWM methods are compared in terms of dc bus utilisation. For the same three-phase sine reference, the PWM method with different zero-sequence signals for the two inverters is found to employ a set of vectors different from the other methods. Both analysis and experimental results show that this method results in lower total harmonic distortion and higher dc bus utilisation than the other two PWM methods.