Figure 1.
Dual transcriptomic strategy for qualitative de novo analysis of cDNA and quantitative analysis of gene expression.
The procedures included sample preparation, cDNA library construction, sequencing, and data analysis that involved de novo assembly, application of BLAST software, gene ontology (GO) annotation, and elucidation of gene expression.
Figure 2.
Synthesis of double-strand cDNA for sequencing.
First strand cDNA synthesis was accomplished by randomized primers, which enabled mRNA to be covered from the poly(A) side to the 5′-side near the start point of transcription.
Figure 3.
Assembly length statistics for marmoset sequences.
Among the 47,883 transcripts, the mean length and SD values were 718 and 457: the longest gene was 7,339 bp in this study.
Table 1.
Summary of sequence assembly.
Table 2.
Annotation statistics of contigs of common marmoset (Callithrix jacchus) transcripts.
Figure 4.
Gene Ontology (GO) classification of marmoset (Callithrix jacchus) sequences based on predicted gene ontology terms in liver, intestine, kidney, and brain from male (A) and female (B) marmosets.
A total of 47,883 contigs with BLAST matches to known proteins were assigned to three main categories: cellular components, molecular function, and biological processes. RNA sequencing was done for mRNAs from liver, intestine, kidney, and brain of three male and three female marmosets. Only the top 100 genes are shown in this GO analysis.
Table 3.
Top 10 transcripts identified or predicted by GO annotation among the most abundant 100 contigs in tissues from male and female common marmosets.
Figure 5.
Expression profiles of marmoset P450 and FMO genes in liver (A), intestine (B), kidney (C), and brain (D) from male and female marmosets.
RNA sequencing was done for mRNAs from liver, intestine, kidney, and brain of three male (black bars) and three female (open bars) marmosets. The mRNA abundance was expressed as the number of fragments. Bars represent the means of pooled tissue samples from three individual marmosets. Known marmoset (Callithrix jacchus) P450 enzymes are shown as Calja P450. The others are named as P450- or FMO-like based on gene annotation.
Table 4.
Representative expression profiles of tissue-specific transporter genes in liver, intestine, kidney, and brain from male and female marmosets.