These sequences were identical to the cDNAs except for three consecutive nucleotides that were absent in two of the three LSUE cDNAs obtained from the EST survey.
While we could not determine the complete sequences for all of the cDNAs, except for Toxoplasma, the remaining sequence information is just as critical to complete.
Compared to de novo fatty acid synthesis, transcript reads for ER-associated lipid enzymes were of low abundance in the non-normalized cDNAs, except for FAD2 Δ12-oleic acid desaturases (1,089 reads) and fatty acid conjugase (284 reads) (Table 4, Figure 6).
To identify the CEACAM gene expression pattern and splice variants, their nucleotide sequences were aligned, and regions of the transmembrane domain-encoding exons with sequence differences were selected to design primers that are specific for each of the CEACAM1-related cDNAs except for CEACAM28 and CEACAM30 which were amplified with a common primer pair (Table 3).
One μg RNA was reverse transcribed with Superscript II RNase H Reverse Transcriptase (Invitrogen) to 20 μl cDNA except for the determination of PEDF transcript.
The sequences of these cDNAs were identical to the canonical FOXO1A cDNA except for the splicing out of 492 bp (extending from nt 2368 285959, FOXO1A-2) and 526 bp (extending from nt 2368 2893, FOXO1A-3)(Fig. 4).
Samples from two cynomolgus monkeys, a 16-year-old female (Philippine origin) and a 15-year-old male (Cambodian-Thai hybrid), were used for the cDNA libraries, except for the liver cDNA library (Qlv).
The reaction mixtures contained 1× Power SYBR Green Master Mix reagent (Applied Biosystems), 250 nM gene-specific primers (except Cs18S, for which 25 nM gene specific-primers were used) and 0.4 µl of the previously synthesized cDNA (except Cs18S for which 0.4 µl of a 25-fold fresh dilution of cDNA was used) in a final volume of 10 µl.
No sequence optimization or alteration was carried out for the cDNAs except to exclude certain restriction enzyme sites from the body of the open reading frame by synonymous codon engineering so as to facilitate cloning into the expression vectors.
Most of the microvillar protein transcripts are abundant in the blood fed cDNA library except for LuloMVP3, which is highly represented in the sugar fed and post blood meal digestion cDNA libraries.
