May 1, 2012

A Member of the Whirly Family is a multifunctional RNA- and DNA- Binding Protein that is Essential for Chloroplast Biogenisis

This study examines the protein ZmWHY1 and its importance in the promotion of atpF intron splicing in vitro. It also disproves the argument that ZmWHY1 only interacts with DNA and shows its interaction with RNA as well. CRS1 is a known agent in the splicing of an atpF intron; however, it is not efficient enough to do the job on its own, other proteins must be involved. Mass spectrometry (a method of identifying chemicals in a substance based on their mass and charge) was used to identify the other proteins involved in the process of splicing an atpF intron. At the conclusion of this process, ZmWHY1 was found to be one of CRS1 protein’s aids in the splicing process. Observations of disruption between ZmWHY1 and CRS1 in the presence of DNAse or RNAse led the researchers to the conclusion that ZmWHY1 interacts with both DNA and RNA. Further investigation showed that the atpF intron RNA was most attracted by the RNA of ZmWHY1.

References:

Prikryl, J., Watkins, K., Frisco, G., Wijk, K., Barkan, A. 2008. Nucleic Acids Research 36:16.

http://nar.oxfordjournals.org/content/36/16/5152.full

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April 5, 2012

atpF intron from 4-12 to 4-22

Sherri Field Gard and Melissa Fry conducted a PCR amplification of extracted orchid DNS’s 4-12 through 4-22 for target region atpF intron. Samples 4-14 and 4-20 both failed to produce a viable yield. All of the other samples produced either a strong or mild success rate. After comparing our samples with the provided molecular mass ruler, we determined that the majority of our samples measured approximately 1,000 bp.

Based on the plastid gene map of Phalaenopsis aphrodite we were able to identify the location of atpF intron. In relation to the gene map, atpF intron is related to genes that code for subunits of ATP synthase complex.

During gel electrophoresis set up, one of our primers, 4-16, was out of order and subsequently was misplaced in the lane order. As seen on the image below, we moved sample 4-16 to the bottom lane to the far right.

Above: Figure 1

LANE NUMBER	DNA CODE	GENUS SPECIES	PCR RESULTS
GEL MARKER
NEGATIVE CONTROL
1	4-12	Chroniochilus virescens	++
2	4-13	Thrixspermum bromeliforme	++
3	4-14	Holcoglossum flavescens	–
4	4-15	Renanthera storei	+
5	4-16	Trichoglottis amesiana	++
6	4-17	Omoea philippinensis	++
7	4-18	Gastrochilus acutifolius	+
8	4-19	Trichoglottis bipunctata	+
9	4-20	Malleola constricta	–
10	4-21	Phalaenopsis lobbii	++
11	4-22	Armordorum siamense	++

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March 2, 2012

Chiloschista segawae

Specimen from which DNA was extracted. Photo by David Jarrell

Chiloschista segawae Masam and Fukuy is a leafless epiphyte, a nonparasitic plant that grows on another plant, commonly found growing on tree trunks in the forests of Taiwan. This particular species grows best in humid, well drained media at warm to cool temperatures. The stem of a Chiloschista segawae is usually hidden by its roots that grow to a length of 35 centimeters long. Its blooming season is in the spring and it produces loose hanging clusters of blooms that measure approximately two inches long. The floral dimensions and arrangement are consistent; however, the floral color pattern can range creating variability concerning the appearance of brown spots versus no brown spots, location of the spots, color intensity or a total yellow appearance.

Chiloschista segawae was named by its authorities, Masam and Fukuy in 1938 and it is a member of the subtribe Aeridinae.