Peran Daya Dukung Wilayah Terhadap Pengembangan USAha Peternakan Sapi Madura

Research conducted on the island of Madura. The aim of the research was analyzed the area-based development of beef cattle in Madura island. Primary research data was sourced from statistics in the Madura district in figures. Data was analyzed using Location Quotient (LQ) method. Data procesing conducted whith spreadsheet from Excel on Microsoft Windows 7. The results showed that the basis for the development of Madura cattle each regency were Pamekasan (sub-district Larangan, Pasean, Batumamar, Palengan, Proppo, Tlanakan, and Pegantenan), Sumenep (sub-district Gayam, Nonggunong and Batuputih), Bangkalan (subdistrict Kokop, Geger, Galis, Tanah Merah, and Blega) and Bangkalan (sub-district Ketapang, Sokobanah, Kedungdung, Sampang, Banyuates, Robatal, and Omben. Conclusion of the research was the development of Madura cattle concentrated in the base region of Madura cattle

Multiple-site fragment deletion, insertion and substitution mutagenesis by modified overlap extension PCR

<p>Introducing various mutations at multiple specific sites within a gene requires multiple steps of DNA manipulation, which is the initial, but limiting step of protein structure–function studies. In the present work, we standardized a simple and fast procedure to perform site-directed mutagenesis, multiple-site fragment deletion, insertion and substitution mutagenesis by a modified version of overlap extension polymerase chain reaction (PCR). In this procedure, target genes divided into several fragments based on the site of mutagenesis are amplified and annealed with their complementary overhanging, followed by extension and amplification to full-length gene with expected mutation(s) by PCR. Vectors inserted with the modified target gene are screened by colony PCR. By using the standardized procedure, we have easily generated single-site mutations, replaced/deleted DNA fragment into/from a target gene and engineered a cysteine-free protein. Practically, the standardized procedure provides an efficient choice for almost all kinds of mutagenesis, especially for multiple-site and large DNA fragment modification mutagenesis. Therefore, this method can be utilized to analyze protein structure and function, to optimize codons of genes for protein expression and to assemble genes of interest.</p

Additional file 1: Table S1. of AFEAP cloning: a precise and efficient method for large DNA sequence assembly

Comparisons of AFEAP cloning with common DNA assembly methods. (DOCX 23 kb

Additional file 6: of AFEAP cloning: a precise and efficient method for large DNA sequence assembly

Supporting Information. (DOCX 32 kb

Additional file 9: Table S5. of AFEAP cloning: a precise and efficient method for large DNA sequence assembly

PCR product reannealing conditions. (DOCX 14 kb

Additional file 4: Figure S2. of AFEAP cloning: a precise and efficient method for large DNA sequence assembly

Sequencing validation of number of fragments characterization. The join sites were shown as S1 to S13, and number of fragments for assembly was shown as 2 + V to 12 + V. The overhang sequences were shown. V: vector backbone. (DOCX 11884 kb

G1 cyclin driven DNA replication

<p>The mitotic cell cycle is driven by Cyclin-Dependent Kinases (CDK). CDK activation requires the binding of activatory subunits termed cyclins. Different waves of cyclins are expressed during the cell cycle, enabling CDKs to trigger phase specific events. For instance, S phase cyclins promote the initiation of DNA replication but not chromosome segregation. There are at least 2 explanations for how such regulation is achieved. According to one of the visions, cyclins confer intrinsic substrate specificity to the CDK catalytic subunit. Alternatively a quantitative model has been proposed, according to which ever-increasing CDK activity is required to trigger cell cycle events from G1 to M. If a quantitative control prevails, then an early cyclin should trigger later cycle events if accumulated at high enough levels at the right time and place. We show here that a G1 phase cyclin bears the potential to trigger DNA replication and promote S and G2 phase specific transcription.</p

Additional file 8: Table S4. of AFEAP cloning: a precise and efficient method for large DNA sequence assembly

PCR thermocycling conditions. (DOCX 13 kb

Additional file 5: Figure S3. of AFEAP cloning: a precise and efficient method for large DNA sequence assembly

Sequencing validation of plasmid sizes characterization. Five join sites are S1, S2, S3, S4, and S5. (a) 11.5 kb plasmid; (b) 19.6 kb plasmid; (c) 28 kb plasmid; (d) 34.6 kb plasmid. The overhang sequences were shown. (DOCX 3815 kb

Additional file 2: Table S2. of AFEAP cloning: a precise and efficient method for large DNA sequence assembly

Primers used in this work. (DOCX 32 kb