English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Global: Zendy Plus annual

Presents the access of premium content as premium feature

Premium Content

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the summarisation as premium feature

Summarisation

Insights

Presents the pdf analysis as premium feature

PDF Analysis

Presents the zaia usage as premium feature

ZAIA

Global: Zendy Tools annual

Global: Zendy Free Plan

Global: Zendy Tools monthly

Global: Zendy Plus monthly

Background  The veined rapa whelk  Rapana venosa  is an important commercial shellfish in China and quantitative real-time PCR (qRT-PCR) has become the standard method to study gene expression in  R. venosa . For accurate and reliable gene expression results, qRT-PCR assays require housekeeping genes as internal controls, which display highly uniform expression in different tissues or stages of development. However, to date no studies have validated housekeeping genes in  R. venosa  for use as internal controls for qRT-PCR.    Methods  In this study, we selected the following 13 candidate genes for suitability as internal controls: elongation factor-1 α  ( EF-1α ),  α -actin ( ACT ), cytochrome c oxidase subunit 1 ( COX1 ), nicotinamide adenine dinucleotide dehydrogenase (ubiquinone) 1 α  subcomplex subunit 7 ( NDUFA7 ), 60S ribosomal protein L5 ( RL5 ), 60S ribosomal protein L28 ( RL28 ), glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ),  β -tubulin ( TUBB ), 40S ribosomal protein S25 ( RS25 ), 40S ribosomal protein S8 ( RS8 ), ubiquitin-conjugating enzyme E2 ( UBE2 ), histone H3 ( HH3 ), and peptidyl-prolyl cis-trans isomerase A ( PPIA ). We measured the expression levels of these 13 candidate internal controls in eight different tissues and twelve larvae developmental stages by qRT-PCR. Further analysis of the expression stability of the tested genes was performed using GeNorm and RefFinder algorithms.    Results  Of the 13 candidate genes tested, we found that  EF-1α  was the most stable internal control gene in almost all adult tissue samples investigated with  RL5  and  RL28  as secondary choices. For the normalization of a single specific tissue, we suggested that  EF-1α  and  NDUFA7  are the best combination in gonad, as well as  COX1  and  RL28  for intestine,  EF-1α  and  RL5  for kidney,  EF-1α  and  COX1  for gill,  EF-1α  and  RL28  for Leiblein and mantle,  EF-1α ,  RL5 , and  NDUFA7  for liver , GAPDH ,  PPIA , and  RL28  for hemocyte. From a developmental perspective, we found that  RL28  was the most stable gene in all developmental stages measured, and  COX1  and  RL5  were appropriate secondary choices. For the specific developmental stage, we recommended the following combination for normalization,  PPIA ,  RS25 , and  RL28  for stage 1,  RL5  and  RL28  for stage 2 and 5,  RL28  and  NDUFA7  for stage 3, and  PPIA  and  TUBB  for stage 4.    Discussion  Our results are instrumental for the selection of appropriately validated housekeeping genes for use as internal controls for gene expression studies in adult tissues or larval development of  R. venosa  in the future.

Selection of housekeeping genes as internal controls for quantitative RT-PCR analysis of the veined rapa whelk (Rapana venosa)