Lagging strand replication of rolling-circle plasmids: Specific recognition of the ssoA-type origins in different gram-positive bacteria

Many bacterial plasmids replicate by a rolling-circle mechanism that involves the generation of single-stranded DNA (ssDNA) intermediates. Replication of the lagging strand of such plasmids initiates from their single strand origin (sso). Many different types of ssos have been identified. One group of ssos, termed ssoA, which have conserved sequence and structural features, function efficiently only in their natural hosts in vivo. To study the host specificity of sso sequences, we have analyzed the functions of two closely related ssoAs belonging to the staphylococcal plasmid pE194 and the streptococcal plasmid pLS1 in Staphylococcus aureus. The pLS1 ssoA functioned poorly in vivo in S. aureus as evidenced by accumulation of high levels of ssDNA but supported efficient replication in vitro in staphylococcal extracts. These results suggest that one or more host factors that are present in sufficient quantities in S. aureus cell-free extracts may be limiting in vivo. Mapping of the initiation points of lagging strand synthesis in vivo and in vitro showed that DNA synthesis initiates from specific sites within the pLS1 ssoA. These results demonstrate that specific initiation of replication can occur from the pLS1 ssoA in S. aureus although it plays a minimal role in lagging strand synthesis in vivo. Therefore, the poor functionality of the pLS1 in vivo in a nonnative host is caused by the low efficiency rather than a lack of specificity of the initiation process. We also have identified ssDNA promoters and mapped the primer RNAs synthesized by the S. aureus and Bacillus subtilis RNA polymerases from the pE194 and pLS1 ssoAs. The S. aureus RNA polymerase bound more efficiently to the native pE194 ssoA as compared with the pLS1 ssoA, suggesting that the strength of RNA polymerase–ssoA interaction may play a major role in the functionality of the ssoA sequences in Gram-positive bacteria.

Drilling at the Sun Flower and Snow Flake claims, Lucky Strike and Peterino claims, and Yellow Circle claim group, San Juan County, Utah /

"January 5, 1955."

The history and records of the Elephant Club; comp. from authentic documents now in possession of the Zoölogical Society.

Added t.-p., engr.

The history and records of the Elephant Club /

Advertisements (28p.) bound in at end and inside front cover.

Midnight talks at the club ...

Mode of access: Internet.

Club papers

[v.23̈ Brown, E.O. De senectute. 1914.--[v.24̈ Dodd, W.E. Robert J. Walker, imperialist. 1914.--[v.25̈ Altrocchi, Rudolph. Gabriele d'Annunzio. 1922

Towards a single crystal Raman spectrum of kaolinite at 77 K

The Raman spectra at 77 K of the hydroxyl stretching of kaolinite were obtained along the three axes perpendicular to the crystal faces. Raman bands were observed at 3616, 3658 and 3677 cm−1 together with a distinct band observed at 3691 cm−1 and a broad profile between 3695 and 3715 cm−1. The band at 3616 cm−1 is assigned to the inner hydroxyl. The bands at 3658 and 3677 cm−1 are attributed to the out-of-phase vibrations of the inner surface hydroxyls. The Raman spectra of the in-phase vibrations of the inner-surface hydroxyl-stretching region are described in terms of transverse and longitudinal optic splitting. The band at 3691 cm−1 is assigned to the transverse optic and the broad profile to the longitudinal optic mode. This splitting remained even at liquid nitrogen temperature. The transverse optic vibration may be curve resolved into two or three bands, which are attributed to different types of hydroxyl groups in the kaolinite.

Story Circle : Digital Storytelling Around the World

Story Circle is the first collection ever devoted to a comprehensive international study of the digital storytelling movement. Exploring subjects of central importance on the emergent and ever-shifting digital landscape-consumer-generated content, memory grids, the digital storytelling youth movement, and micro-documentary- Story Circle pinpoints who is telling what stories, where, on what terms, and what they look and sound like.

K-tree : large scale document clustering

We introduce K-tree in an information retrieval context. It is an efficient approximation of the k-means clustering algorithm. Unlike k-means it forms a hierarchy of clusters. It has been extended to address issues with sparse representations. We compare performance and quality to CLUTO using document collections. The K-tree has a low time complexity that is suitable for large document collections. This tree structure allows for efficient disk based implementations where space requirements exceed that of main memory.

Digital storytelling as participatory public history in Australia

This chapter discusses digital storytelling as a methodology for participatory public history through a detailed reflection on an applied research project that integrated both public history and digital storytelling in the context of a new master-planned urban development: the Kelvin Grove Urban Village Sharing Stories project.

The knowledge advantage (K-Adv): unleashing creativity and innovation

The K-Adv has been developed around the concept that it comprises an ICT enabling infrastructure that encompasses ICT hardware and software infrastructure facilities together with an enabling ICT support system; a leadership infrastructure support system that provides the vision for its implementation and the realisation capacity for the vision to be realised; and the necessary people infrastructure that includes the people capabilities and capacities supported by organisational processes that facilitates this resource to be mobilised.