Construction of bovine adenovirus type 3 E1 and E3, substitution plasmids and the sequencing analysis of DNA pol and pTP /

The relative ease to concentrate and purify adenoviruses, their well characterized mid-sized genome, and the ability to delete non-essential regions from their genome to accommodate foreign gene, made adenoviruses a suitable candidate for the construction of vectors. The use of adenoviral vectors in gene therapy, vaccination, and as a general vector system for expressing foreign genes have been documented for some time. In this study, the objective was to rescue a BAV3 E1 or E3 recombinant vector carrying the kanamycin resistant gene, a dominant selectable marker with useful applications in studying vectored gene expression in mammalian cells. To accomplish the objective of this study, more information about BAV3 DNA sequences was required in order to make the manipulation of the virus genome accessible. Therefore, sequencing of the BAV3 genome from 1 1 .7% to 30.8% was carried out. Analysis of the determined sequences revealed the primary structure of important viral gene products coded by E2 including BAV3 DNA pol and precursor to terminal protein. Comparative analysis of these proteins with their counterparts from human and non human adenoviruses revealed important insights as to the evolutionary lineage of BAV3. In order to insert the kanamycin resistance gene in either E1 or E3, it was necessary to delete BAV3 sequences to accommodate the foreign gene so as not to exceed the limit of the packaging capacity of the virus. To construct a recombinant BAV3 in which a foreign gene was inserted in the deleted E1 region, an E1 shuttle vector was constructed. This involved the deletion from the viral sequences a region between 1.3% to 9% and inserting the kanamycin resistance gene to replace the deletion. The E1 shuttle vector contained the left (0%- 53.9%) segment of the genome and was expected to generate BAV3 recombinants that can be grown and propagated in cells that can complement the missing E1 functions. To construct a similar shuttle vector for E3 deletion, DNA sequences extending from 78.9% to 82.5% (1281 bp) were deleted from within the E3 region that had been cloned into a plasmid vector. The deleted region corresponds to those that have been shown to be non-essential for viral replication in cell culture. The resulting plasmid was used to construct another recombinant plasmid with BAV3 DNA sequences extending from 37.1% to 100% and with a deletion of E3 sequences that were replaced by kanamycin resistance gene. This shuttle plasmid was used in cotransfections with digested viral DNA in an attempt to rescue a recombinant BAV3 carrying the kanamycin resistance gene to replace the deleted E3. In spite of repeated attempts of transfection, El or E3 recombinant BAV3 were not isolated. It seems that other approaches should be applied to make a final conclusion on BAV3 infectivity.

A. S. Whiting Manufacturing Company Limited, minute book, 1872-1909.

The Cedar Dale Scythe Works was the second manufacturing company that A.S. Whiting had established in Oshawa, the first being the Oshawa Manufacturing Co. in 1852. The Oshawa Manufacturing Co. was eventually taken over by the Joseph Hall Works in 1857. In 1862, the Cedar Dale Works was built after being in a rented space in the Hall Works for two years, building scythes and hoes. With the building of the firm, the village of Cedar Dale was established. In 1867, the firm became Whiting and Cowan when John Cowan bought into the company. After the death of Whiting in 1867, his son-in-law, R.S. Hamlin headed the company. By 1872, it became the A.S. Whiting Manufacturing Co. when Cowan withdrew from the business. Before Whiting’s death, the company had been profitable but due to new machinery being developed, hand tools were becoming obsolete and the business only lasted for a few more years (source: Oshawa Community Museum and Archives Web site).

Proceedings : Grand Lodge, A.F. & A.M. of Canada in the Province of Ontario - 1872

On spine : The Grand Lodge of Canada in the Province of Ontario.

Wakeman Burritt fonds, 1807-1814

Wakeman Burritt (1785-1847) was a merchant and ship owner who conducted his business in New York, New Orleans, LA, Charleston, SC, and the West Indies. Despite the difficulties that the Embargo Act of 1807 and the War of 1812 posed to trade and commerce, Burritt managed to run a successful business, dealing in commodities such as foodstuffs, cloth, cotton, and soap. He owned three ships, the Brig Cannon, Brig Eunice and Brig Sampson. In 1809, he married Grace Burr (1787-1874). Together they had at least one child, Francis Burritt (1811-1861).

Photograph - Agnes Ethelwyn Wetherald, 1872.

Photograph of Ethelwyn Wetherald at age 15 in the year 1872. The same year her youngest sibling, Herbert, was born.

Proceedings of the Grand Chapter of Royal Arch Masons of Canada at the annual convocation, 1872

Annual Convocation proceedings for the year 1872. The title varies slightly and convocation is held at different cities or towns in Canada. Fifteenth annual convocation.

Photograph - Isaiah Hansler, 1872

A photograph of Isaiah Hansler (1851-1931) circa 1872.

Copy of poster advertising “Grand Buffalo Hunt at Niagara Falls” with Bill Hickok, August, 1872

James Butler Hickok (1837-1876), also known as “Wild Bill” Hickok, was an American gun-fighter, scout and spy. He was involved in altercations with others while working for the famous express company Russell, Majors and Waddell (in 1861), and later while working as a wagon master, scout, and spy for the Union forces during the Civil War. These altercations resulted in the deaths of 4 people, but Hickok was acquitted in all cases. An embellished article written about him in Harper’s magazine helped contribute to his reputation as a western hero. He served as a deputy U.S. marshal and sheriff in Kansas in the late 1860’s, helping to bring law and order to a previously lawless area. He gambled considerably, and during a card game on August 2, 1876, was shot and killed. The cards he was holding (two aces, two eights, and a jack) became known as the "dead man's hand."

License no. 144 of season 1872/73 made out to S.D. Woodruff for 36 square miles in berth no. 192

License no. 144 of season 1872/73 made out to S.D. Woodruff for 36 square miles in berth no. 192. This document is slightly torn and stained along the right hand side. This does not affect the text, April 7, 1873.